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Molecular and morphological changes in the cat lateral geniculate nucleus and visual cortex induced by visual deprivation are revealed by monoclonal antibodies Cat-304 and Cat-301.

Monoclonal antibody Cat-301 recognizes a surface-associated proteoglycan on subsets of neurons in the mammalian CNS (Hockfield and McKay, 1983). The expression of Cat-301 immunoreactivity on Y cells in the cat LGN is sharply reduced by early visual deprivation (Sur et al., 1988). We employed an immunosuppression strategy (Hockfield, 1987) to further study alterations in the expression of experience-dependent molecules. Newborn BALB/c mice were injected with LGN from dark-reared cats to induce a suppression of the immune response to antigens expressed in visually deprived animals. These mice were then immunized with LGN from normal cats to elicit an immune response to antigens with an expression dependent on normal early visual experience. This strategy permitted the generation of monoclonal antibody Cat-304, which recognizes a surface-associated antigen on neuronal cell bodies and proximal dendrites, and which appears histologically identical to Cat-301. Further analyses show that Cat-304 and Cat-301 recognize different epitopes on the same 680-kDa chondroitin sulfate proteoglycan. We examined the effects of early visual deprivation on Cat-304 immunoreactivity in the LGN and visual cortex of cats. In LGN from normal cats, Cat-304 labels neurons in layers A, A1, and C, in interlaminar zones, and in the medial interlaminar nucleus. In LGN from dark-reared cats, the number of antibody-positive neurons is markedly reduced, and the cross-sectional area of the remaining positive neurons is smaller than normal. In cortical area 17 of normally reared cats, Cat 304-positive neurons are densely distributed in 2 bands, in layers IV and V/VI. Labeled neurons are also present in layers II and III. In area 17 of dark-reared cats, the number of antibody-positive neurons is reduced. The reduction in the number of labeled neurons is most pronounced in layers II/III and V/VI. Antibody-positive neurons are smaller in all cortical layers of dark-reared cats. The changes in the expression of Cat-301 immunoreactivity in dark-reared visual cortex and LGN are identical to those of Cat-304. The laminar differences in the effect of dark rearing on Cat-301 and Cat-304 expression in the visual cortex provides support for the suggestion that layer IV of cortical area 17 may be less susceptible to prolongation of plasticity by dark rearing than layers II/III and V/VI. Further, the biochemical and histological studies reported provide evidence that early visual experience regulates protein expression in the cat LGN and visual cortex.

Animals

Stereopsis in normal domestic cat, Siamese cat, and cat raised with alternating monocular occlusion.

Normal cats, cats raised with alternating monocular occluders, and Siamese cats were tested for stereoscopic vision using a shadow-casting technique that produced line stereograms. Human subjects were tested with the same apparatus. 1. Normal cats had stereoscopic vision, but AO and Siamese cats did not. 2. Normal AO cats, and Siamese cats all had equal visual activity (about 6 feet of arc) and all could make accurate judgments about the depth of real objects. 3. Normal cats could fuse crossed disparities as great as 50 feet of arc and uncrossed disparities as great as 30 feet of arc. 4. Normal cats could make stereoscopic discriminations with stimulus disparities greater than 1 degree, even though they could not fuse disparities of this magnitude. 5. Both the AO and the Siamese cats exhibited a convergent squint. 6. Human subjects viewing identical stimuli could not fuse stimuli having disparities greater than 10 feet, although they could make depth judgments for crossed disparities as great as 5 degrees. 7. These results imply that animals without binocular cells in area 17 do not have stereoscopic vision, but do not determine if disparity-selective cells in the visual cortex are responsible for stereopsis.

Animals

Myocardial taurine concentrations in cats with cardiac disease and in healthy cats fed taurine-modified diets.

Myocardial taurine concentrations were measured in cats with cardiac disease and in healthy cats fed diets with various concentrations of taurine. Group 1 was composed of 26 cats with 3 categories of naturally developing cardiac disease: dilatative cardiomyopathy (group 1A), 10 cats; hypertrophic cardiomyopathy (group 1B), 9 cats; and volume overload (group 1C), 7 cats. These cats had been fed various commercial diets. Group 2 was composed of 40 healthy cats that had been fed diets varying in taurine concentration (0 to 1% taurine) for at least 2 years. Mean myocardial taurine concentrations did not differ significantly between group-1 cats with dilatative cardiomyopathy and those with hypertrophic cardiomyopathy or volume overload. Cats in group 1A had a mean myocardial taurine concentration 3 times higher than healthy cats fed a taurine-free diet (P less than 0.002). Mean myocardial taurine concentrations did not differ significantly between group-1A cats and healthy cats fed a diet containing 0.02% taurine; group-1A cats had significantly lower mean myocardial taurine concentrations than did healthy cats fed a synthetic diet containing 0.05 or 1.0% taurine (P less than 0.001). Acute oral administration of taurine in 5 group-1A cats appeared to increase mean myocardial taurine concentrations, compared with similar cats not given taurine during treatment for cardiac failure. In group-2 cats, mean myocardial taurine concentrations increased directly with percentage of dietary taurine.

Animals

Pharmacokinetics of methimazole in normal cats and cats with hyperthyroidism.

The intravenous and oral disposition of the antithyroid drug methimazole was determined in 10 clinically normal cats and nine cats with naturally occurring hyperthyroidism. After intravenous administration of 5 mg methimazole, the mean residence time was significantly (P less than 0.05) shorter in the cats with hyperthyroidism than in the normal cats, but there was no significant difference between the mean values for total body clearance (CL), steady state volume of distribution (Vdss), terminal elimination rate constant (ke), or serum terminal half-life (t1/2) in the two groups of cats. After oral administration, the mean bioavailability of methimazole was high in both the normal cats (77.6 per cent) and cats with hyperthyroidism (79.5 per cent). The values for mean residence time, ke and serum terminal t1/2 after oral dosing were significantly shorter in the cats with hyperthyroidism than in the normal cats. However, after oral administration of methimazole there were no significant differences between the mean values for CL, Vdss, bioavailability and maximum serum concentrations or the time for maximal concentrations to be reached in the two groups of cats. Overall, most pharmacokinetic parameters for methimazole were not altered by the hyperthyroid state. However, the cats with hyperthyroidism did show a trend toward faster elimination of the drug compared with the normal cats, similar to what has been previously described for the antithyroid drug propylthiouracil in cats. These results also indicate that methimazole is well absorbed when administered orally and has a higher bioavailability than that of propylthiouracil in cats with hyperthyroidism.(ABSTRACT TRUNCATED AT 250 WORDS)

Absorption

Plasma taurine concentrations and M-mode echocardiographic measures in healthy cats and in cats with dilated cardiomyopathy.

M-mode echocardiography was completed and plasma taurine concentrations were determined in 79 healthy cats and 77 cats with dilated cardiomyopathy (DCM). In healthy cats, a relationship was not observed between plasma taurine concentrations and any M-mode echocardiographic measurement. End-systolic and end-diastolic cardiac chamber dimensions were larger; wall thickness measures were smaller; and calculations of fractional shortening were less in cats with DCM than in healthy cats. Plasma taurine concentrations less than 30 nmol/mL were detected in 7/79 healthy cats and in 52/77 cats with DCM. Of the 52 cats with DCM and an initial plasma taurine concentration less than 30 nmol/mL, 23 died or were euthanized during the first post-treatment week, 7 were lost to further study, and 22 improved after taurine supplementation. Of the 25 cats with DCM and an initial plasma taurine concentration greater than or equal to 30 nmol/mL, 9 died or were euthanatized during the first post-treatment week, and 9 were lost to further study. Two cats did not improve, of which one died and one was euthanatized 4 to 8 weeks after initiation of taurine supplementation. Five cats with a plasma taurine concentration greater than or equal to 30 nmol/mL improved after taurine supplementation. Myocardial function subsequently deteriorated in three of these cats. Two of the three cats had signs of congestive heart failure redevelop.

Animals

Sperm capacitation in the domestic cat (Felis catus) and leopard cat (Felis bengalensis) as studied with a salt-stored zona pellucida penetration assay.

The ability of domestic cat or leopard cat spermatozoa to penetrate zonae pellucidae (ZP) of salt-stored, domestic cat oocytes was examined as an assay for sperm capacitation. Ovarian oocytes were recovered after ovariectomy and matured in vitro for 18-36 h. Following removal of cumulus cells, the oocytes were used fresh, or stored (4 degrees C, 0.5-24 weeks) in a HEPES-buffered hypertonic salt solution. Electroejaculated, washed sperm (2-4 x 10(6) sperm/ml) were preincubated for 1.0 h (38 degrees C, 5% CO2 in air) and then co-incubated (2 x 10(5) sperm/ml) with fresh or stored oocytes for 6.0 h. Gametes were incubated in a protein-free, modified Tyrode's solution (TLP-PVA) or in the same medium containing 4.0 mg/ml bovine serum albumin (BSA; TALP-PVA). Treatments were compared for percentage ZP penetration (defined as sperm heads reaching more than halfway through the ZP) as an index of sperm capacitation. In both the domestic cat and leopard cat, there was no difference (P greater than 0.05) in sperm penetration of fresh ZP (domestic cat, 42.5 +/- 5.4%; leopard cat, 38.6 +/- 2.8%) or stored ZP (domestic cat, 32.4 +/- 4.2%; leopard cat, 27.6 +/- 2.3%). Sperm incubated in protein-free medium (TLP-PVA) were less capable (P less than 0.05) of ZP penetration (domestic cat, 14.6 +/- 5.9%; leopard cat, 7.9 +/- 3.0%) than sperm incubated in medium TALP-PVA containing BSA (domestic cat, 60.3 +/- 5.9%; leopard cat, 58.4 +/- 3.0%).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Role of thyroid hormones in an abnormal grooming behavior in thyroidectomized cats and cats with pontile lesions.

Thyroidectomized cats and cats with pontile lesions exhibit the same abnormal grooming behavior. Investigations were conducted to determine whether the abnormal grooming behavior in cats with pontile lesions was attributable to a hypothyroid condition. Administration of thyroid hormones abolished the abnormal behavior of thyroidectomized cats but was without effect in cats with pontile lesions. Studies of brain and blood levels of thyroid hormones in cats with pontile lesions failed to detect any differences from normal cats, and normal effects of thyroid hormones on evoked potentials to light flashes and to clicks were obtained in cats with pontile lesions. It was concluded that any thyroid dysfunction that may exist in the cats with pontile lesions was not involved in the genesis of the abnormal grooming behavior. Because the abnormal grooming behavior fluctuates rhythmically during the year, an endocrine hypothesis was entertained as the source of the fluctuations; endocrine concomitants of the abnormal behavior were investigated with longitudinal studies. The urinary excretion of 11-hydroxycorticoids exhibited a significant negative correlation with the abnormal behavior. When compared with that of normal cats, the plasma level of 11-hydroxycorticoids of thyroidectomized cats was significantly decreased. Intramuscular injections of 11-hydroxycorticoids abolished the abnormal behavior in both thyroidectomized cats and cats with pontile lesions. These results considered together with previous findings indicate that thyroidectomy and pontile lesions induce similar changes in 11-hydroxycorticoid and serotonergic functions-changes that are critical features of the physiological bases of the abnormal grooming behavior in both groups.

11-Hydroxycorticosteroids

Reported cat bites in Dallas: characteristics of the cats, the victims, and the attack events.

Associated with the increased popularity of cats as pets in American households has been an increase in the number of cat bites reported to health departments. Bite reports from Dallas, TX, for 1985 were analyzed for different aspects of the cat bite event, including characteristics of the cats, the people bitten, the wounds, and the attack events. Cat bites and scratches constituted 25 percent of the 2,494 reported animal bites. Biting cats were typically stray females. People 21 to 35 years old were bitten in numbers disproportionate to their numbers in the Dallas population. Females were the victims of a majority of bites. Although wounds were typically described as "scratches" of a hand or finger, 80 percent of all victims sought some form of treatment for their wound. The highest proportion of bites occurred from May through August from 9 am through 12 noon; unowned cats accounted for most wounds. Cat bit events may be explained by frequency of contact, that is, women prefer cats as pets; activities that bring people and cats into contact; and reaching toward cats to feed or pet them. More specific information on the causes of cat bites will enable educational programs to be established so that the rate of cat bites can be decreased at the community level.

Adolescent

Correlation of urine protein/creatinine ratio and twenty-four-hour urinary protein excretion in normal cats and cats with surgically induced chronic renal failure.

Urine protein/creatinine (UP/C) ratios and 24-hour urinary protein excretion were compared in clinically normal cats and cats with surgically induced chronic renal failure (CRF). Mean 24-hour urinary protein excretion in 30 clinically normal cats fed a 28% protein diet (dry weight basis) was 4.93 mg/kg/24-hour (SD = 1.34) with a range of 2.99 to 8.88. Mean UP/C ratio in these cats was 0.134 (SD = 0.037) with a range of 0.073 to 0.239. Mean 24-hour urinary protein excretion in CRF cats was 10.49 mg/kg/24-hour (SD = 11.28) with a range of 2.16 to 62.93. Mean UP/C ratio in the CRF cats was 0.359 (SD = 0.374) with a range of 0.061 to 1.916. Linear regression showed high correlation (R2 = 0.973, P less than 0.001) between 24-hour urinary protein excretion and UP/C ratio in clinically normal cats and cats with surgically induced chronic renal failure. The regression equation for 24-hour urinary protein excretion versus UP/C ratio was: 24-hour urinary protein excretion = 29.39 (UP/C) + 0.18. Results of this study indicate that UP/C ratios are a valid estimate of 24-hour urinary protein excretion in clinically normal and CRF cats. Dietary protein intake significantly affected UP/C ratios in clinically normal cats and cats with surgically induced CRF. Therefore, the influence of dietary protein should be considered when interpreting UP/C ratios.

Animals

A survey of helminths in domestic cats in the Pretoria area of Transvaal, Republic of South Africa. Part 1: The prevalence and comparison of burdens of helminths in adult and juvenile cats.

The helminths found in 1,502 necropsied cats were examined. The findings indicated that 65% of the cats were infested. The most prevalent helminths encountered were Ancylostoma tubaeforme (41%), Ancylostoma braziliense (25%), Dipylidium caninum (23%), Toxocara cati (11%), Taenia taeniaeformis (7.7%), Ancylostoma caninum (3.3%), Joyeuxiella fuhrmanni (2.5%), Ancylostoma ceylanicum (1.4%), and Physaloptera praeputialis (1.3%). The following helminths were recorded in fewer than 1% of the cats: Centrorhynchus spp., Pterygodermatites spp., Toxocara canis, Aleurostrongylus abstrusus and Vogeloides spp., This is the first record of the lungworms, A. abstrusus and Vogeloides spp. in cats in the Republic of South Africa. The helminths in adult and juvenile cats are compared. Of the cestodes, Joyeuxiella fuhrmanni (4%) and Taenia taeniaeformis (12%) are more prevalent in adult cats. Dipylidium caninum is marginally more common in adults (24%), and is the most common helminth in juvenile cats, being present in 21% of cases. More adult cats have Ancylostoma tubaeforme (58%), Ancylostoma braziliense (36%), Ancylostoma ceylanicum (2.4%) and Physaloptera praeputialis (1.9%) than do juvenile cats. However, juvenile cats were found to harbour more Toxocara cati (41%), Toxocara canis (0.5%) and Ancylostoma caninum (5.2%). Juvenile cats harbour appreciable numbers of both cestodes and nematodes, thus any treatment should be aimed at the elimination of both, with a broad spectrum anthelmintic.

Age Factors

Cat-301 antibody selectively labels neurons in the Y-innervated laminae of the cat superior colliculus.

Cat-301 is a monoclonal antibody which recognizes a cell surface associated antigen of selected neurons in the central nervous system (CNS). In the visual system, cat-301 selectively labels Y-like cells in several visual structures, including portions of the lateral geniculate nucleus complex and visual cortex. The cat superior colliculus (SC) also receives Y input and contains cells driven by Y input which are selectively distributed in the deep superficial gray and deeper laminae. If cat-301 is selective to the Y-cell system in SC, labeled cells should be restricted to those laminae. To test this hypothesis, we have examined quantitatively the laminar distribution, percentage, size, and morphology of cells in SC labeled by the cat-301 antibody. Cat-301 labeled a variety of cells in the cat SC. Labeled cells were found within the deep portion of the superficial gray layer (6.6%), optic layer (27.6%), intermediate gray layer (26.9%), and the deep gray and white layers (38.5%). By contrast, only 2 of 667 labeled cells (0.3%) were found within that part of the upper superficial gray layer innervated exclusively by W input and thought to contain only W-driven cells. When considered as a percentage of the total cell population, cat-301 labeled cells represented less than 3% of cells in the superficial gray layer and approximately 15% in the deeper layers. Neurons labeled by cat-301 were all of medium to large size (mean average diameter = 33.3 microns; range = 15-84 microns) and included vertical fusiform and stellate cells in the upper layers and the very large neurons found in the intermediate gray and deeper layers. These results provide further evidence that the cat-301 antibody selectively recognizes the Y channel of the cat visual system.

Animals

Comparison of fractional excretion and 24-hour urinary excretion of sodium and potassium in clinically normal cats and cats with induced chronic renal failure.

The influence of induced chronic renal failure on 24-hour urinary excretion and fractional excretion of sodium and potassium was studied in cats. Induction of chronic renal failure significantly increased fractional excretion of potassium (P less than 0.0001) and sodium (P less than 0.05); however, 24-hour urinary excretion of sodium and potassium decreased slightly following induction of chronic renal failure. Fractional excretion and 24-hour urinary excretion of sodium and potassium were compared by linear regression in clinically normal cats, cats with chronic renal failure, and clinically normal and affected cats combined. In clinically normal cats, linear regression revealed only moderate correlation between fractional excretion and 24-hour urinary excretion for sodium and potassium. Linear regression of these same relationships in cats with chronic renal failure, and in clinically normal cats and cats with chronic renal failure combined, indicated low correlation. Fractional excretions of sodium and potassium were not reliable indicators of 24-hour urinary excretion of these electrolytes in cats with chronic renal failure or unknown glomerular filtration rate. Fractional excretion of potassium and sodium correlated only moderately with 24-hour urinary excretion in clinically normal cats.

Animals

Ejaculate-hormonal traits in the leopard cat (Felis bengalensis) and sperm function as measured by in vitro penetration of zona-free hamster ova and zona-intact domestic cat oocytes.

Electroejaculate traits and circulating follicle-stimulating hormone (FSH), luteinizing hormone (LH), and testosterone concentrations were analyzed in adult leopard cats (Felis bengalensis), a rare felid species indigenous to east Asia. The ability of leopard cat sperm to bind and penetrate zona-free hamster ova and zona-intact domestic cat oocytes in vitro was examined as a means of testing sperm function. The influence of culture media [Biggers, Whitten, Whittingham (BWW) vs. modified Krebs Ringer bicarbonate (mKRB)], seminal plasma removal, and swim-up separation on sperm motility, sperm morphology, and oocyte penetration also were assessed. Sperm treatments included dilution of raw semen (DR), ejaculate centrifugation, and either resuspension (NS) or swim-up processing (SU). The percentage of oocytes penetrated (penetration rate) and the number of penetrated sperm/oocyte (penetration index) were determined. Ejaculates from each male consisted of at least a 50% sperm motility rating, and hormone concentrations in individual males were unrelated to any ejaculate trait measured concurrently on the same day. The SU technique improved (P less than 0.05) percent sperm motility and the proportion of structurally normal sperm compared to DR and NS treatments. Leopard cat spermatozoa were capable of binding to and penetrating hamster ova and domestic cat oocytes; however, penetration was influenced by culture medium and seminal processing. In the hamster assay, a higher (P less than 0.05) penetration rate and penetration index were achieved when mKRB was used for gamete incubation instead of BWW. NS processing also increased (P less than 0.05) overall penetration compared to DR and SU. In the cat oocyte assay, zona penetration rate was similar (P greater than 0.05) in the DR, NS, and SU aliquots; however, the zona penetration index was increased (P less than 0.05) by the NS compared to the DR and SU treatments. This study 1) provides baseline ejaculate and endocrine norms for the leopard cat, 2) demonstrates that leopard cat sperm undergo nuclear decondensation in hamster ova and penetrate zona-intact domestic cat oocytes, 3) indicates that seminal plasma removal enhances leopard cat sperm fertilizing ability and ovum penetration, and 4) suggests that heterologous oocyte penetration is effective for assessing factors influencing fertilization and sperm function in this nondomestic felid.

Animals

Airborne cat allergen (Fel d I). Environmental control with the cat in situ.

In a house with a cat furnishings, air-exchange rate, and the cat are all thought to influence airborne cat allergen. We carried out experiments using two separate rooms, modifying the environment, applying different cleaning techniques, and washing the cat, to analyze these sources and to design methods of reducing airborne allergen. Airborne measurements were made with a cascade impactor and a two-site monoclonal antibody-based immunometric assay for cat allergen Fel d I. Within 30 min of entering a 30-m3 clean room the cat itself was found to increase airborne Fel d I by 30 to 90 ng/m3. Following serial weekly washing of the cat this increase was reduced to less than or equal to 7 ng/m3, with a more marked fall in small particles (less than or equal to 2.5 microns diameter) from 9.5 to less than or equal to 0.4 ng/m3. To study the influence of the room design we kept the cat in a room of 33 m3 for 20 h/day and modified the room. This room was studied with or without furnishings and with air-exchange rates of 0.2 or 2.4 air changes per hour. Both low ventilation rate and furnishings increased the level of Fel d I measured 1 h after the cat was removed. However, the most striking finding was that the carpet accumulates cat allergen at approximately 100 times the level for a polished floor, that is, approximately 100 micrograms/day Fel d I compared with approximately 0.5 micrograms/day Fel d I.(ABSTRACT TRUNCATED AT 250 WORDS)

Air

Erythrocyte protoporphyrin concentrations in clinically normal cats and cats with lead toxicity.

Erythrocyte protoporphyrin (EPP) and blood lead concentrations were determined in 91 clinically healthy cats living in the inner suburban area of Sydney, Australia. The mean EPP concentration was 223.4 +/- 186.1 micrograms litre-1 whole blood and the mean blood lead concentration 0.62 +/- 0.25 mumol litre-1. EPP concentrations were also monitored in three cats with confirmed lead toxicity--at the time of diagnosis and one week and one month after chelation therapy with calcium EDTA. EPP concentrations were elevated in two cats and within the normal range in the third cat at the time of diagnosis. EPP concentration were higher in two cats one week after treatment than at the time of diagnosis. One month after chelation therapy, EPP concentrations were normal in two cats but still substantially elevated in the third cat although its blood lead concentration had returned to normal and all clinical signs of lead toxicity had resolved. It was determined that the predominant form of protoporphyrin present in cats with lead toxicity was zinc protoporphyrin. The EPP assay may have limited value in the diagnosis of acute lead toxicity and in monitoring the success of chelation therapy in cats.

Animals

Effects of cat exposure and cat odors on subsequent amphetamine-induced stereotypy.

The effect of exposure to a cat, as a predatory stressor, was examined in male and female rats during subsequent tests of amphetamine-induced stereotypy in which cat odors were present or absent. Rats in Group C/O were given a 15-min exposure session to a male cat while they were protected in a wire cage. They were then given an IP injection of d-amphetamine (1 mg/kg) and tested 30 min later for stereotypy in the presence of cat odors (soiled cat litter). Rats in Group NC/O were given a no-cat-exposure control session, and amphetamine tested with cat odors. Groups C/NO and NC/NO were both tested without cat odors (fresh litter), with the former group having been previously exposed to a cat. During the 90-min test sessions, female rats showed significantly more stereotypy than males. More importantly, the male subjects in group C/O exhibited significantly more stereotypy than the males in the other groups, and group NC/NO males showed the least amount of stereotypy. These findings clearly indicate that amphetamine reactivity is influenced by prior exposure to a predator, the presence of predatory odors during testing, and the subject's sex.

Animals

Mapping of cat albumin using monoclonal antibodies: identification of determinants common to cat and dog.

Cat and dog albumins from commercial extracts were used to produce monoclonal antibodies (MoAb). Anti-cat albumin MoAb recognized both cat and dog albumin equally, as did anti-dog albumin MoAb; this confirms cross-reactivity between cat and dog. The MoAb were separated into two groups according to their epitopic specificity; they recognized two overlapping epitopes of cat albumin. Furthermore, by competitive inhibition of radio-allergosorbent test (RAST), it was shown that one MoAb group inhibited significantly the binding of human IgE antibodies (from a pool of 13 patients allergic to both cats and dogs) to insolubilized cat or dog extracts. These observations suggest that murine anti-cat or anti-dog MoAb and human IgE antibodies recognize identical or closely related determinants on cat and dog albumin.

Albumins

Fiber type- and position-dependent expression of a myosin light chain-CAT transgene detected with a novel histochemical stain for CAT.

We recently generated and characterized transgenic mice in which regulatory sequences from a myosin light chain gene (MLC1f/3f) are linked to the chloramphenicol acetyltransferase (CAT) gene. Transgene expression in these mice is specific to skeletal muscle and graded along the rostrocaudal axis: adult muscles derived from successively more caudal somites express successively higher levels of CAT. To investigate the cellular basis of these patterns of expression, we developed and used a histochemical stain that allows detection of CAT in individual cells. Our main results are as follows: (a) Within muscles, CAT is detected only in muscle fibers and not in associated connective tissue, blood vessels, or nerves. Thus, the tissue specificity of transgene expression observed by biochemical assay reflects a cell-type specificity demonstrable histochemically. (b) Within individual muscles, CAT levels vary with fiber type. Like the endogenous MLC1f/3f gene, the transgene is expressed at higher levels in fast-twitch (type II) than in slow-twitch (type I) muscle fibers. In addition, CAT levels vary among type II fiber subtypes, in the order IIB greater than IIX greater than IIA. (c) Among muscles that are similar in fiber type composition, the average level of CAT per fiber varies with rostrocaudal position. This position-dependent variation in CAT level is apparent even when fibers of a single type are compared. From these results, we conclude that fiber type and position affect CAT expression independently. We therefore infer the existence of separate fiber type-specific and positionally graded transcriptional regulators that act together to determine levels of transgene expression.

Animals