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H Merchant

Publications and source records attributed to H Merchant.

At least 37 records · Page 2Linked to original sources

Categorization of somaesthetic stimuli: sensorimotor performance and neuronal activity in primary somatic sensory cortex of awake monkeys.

We used psychometric techniques to study the sensorimotor performance of four monkeys trained to classify the speed of moving tactile stimuli. Animals performed the task by pressing one of two target switches to indicate whether the speed of probe movement across the glabrous skin of the hand was low or high. Psychometric curves indicated that animals classified the stimulus speeds irrespective of which finger was stimulated, traverse distance and direction. The mean values of the reaction (RT) and movement (MT) times during the correct categorization of low and high stimulus speeds were similar. However, a slight increase was detected in the mean values of the RT during the incorrect categorization but not in the MT. During the task, activity of single neurones (n = 45) was recorded in primary somatic sensory (SI) cortex. The results indicate that a class of neurones (n = 12) of SI cortex increased their impulse rates as a function of the stimulus speeds. However, the magnitude of their responses was similar during the correct and incorrect categorizations of stimuli. The same neurones also responded when the same set of stimuli used in the categorization task were delivered passively. Neurones of SI cortex responded with a latency of 25.8 +/- 0.6 ms (+/- s.e.m.) relative to the beginning of the moving tactile stimuli during the categorization task. The same neurones (n = 17) also responded with a similar latency (24.6 +/- 4.0 ms) when the stimuli were delivered passively. These results may suggest that, although this evoked neuronal activity may be important for the perception of the moving tactile stimuli, more central structures associated with SI cortex may determine the performance of this learned somaesthetic task.

Animals↗

Neuronal activity of primate putamen during categorical perception of somaesthetic stimuli.

We have studied neuronal activity in the putamen of two monkeys trained to discriminate the speed of moving tactile stimuli. Animals pressed one of two target switches to indicate whether the speed of the probe across the skin was low or high. The activity of single neurones was recorded in the putamen ipsilateral to the glabrous skin of the stimulated hand and contralateral to the responding arm. During the task, we recorded neurones in the putamen that showed responses confined exclusively to the stimulus period of all speeds. A second class of putamen neurones responded during the stimulus period but continued discharging during the reaction and movement time period. None of these two classes of putamen neurones discharged when the same set of stimuli were delivered passively. A third class of putamen neurones responded differentially in the discrimination task and predicted whether the speed of the stimulus was low or high. A number of these neurones, which responded differentially during the categorization task, were tested in a light instruction task. This tested the possibility that these differential responses were associated with the intention to move the arm to one of the two target switches. Few neurones responded in this situation. These results indicate that the putamen, in addition to its role in motor regulation, is also involved in higher order aspects of sensory-motor behaviour and in the sensory decision process in this learned somaesthetic task.

Animals↗

Pituitary-ovarian dysfunction in rats with induced nephrotic syndrome.

The reproductive hormonal profile was evaluated in female rats with experimental nephrotic syndrome induced with a single subcutaneous dose of puromycin aminonucleoside (PAN). Serum concentrations of luteinizing hormone (LH), follicle-stimulating hormone (FSH), progesterone (P4), testosterone and 17 beta-estradiol (E2) were determined sequentially in control and experimental groups on days 1, 3, 7 and 10 after PAN administration. Prolactin levels were also assessed on day 10. In both groups, vaginal smears were taken daily throughout the study to evaluate cyclic histological changes. At the end of the experiment the histological appearance of the ovaries was evaluated by light microscopy. Nephrotic rats had a rapid loss of the estrous cycle starting on day 4, which set them at diestrus. At the same time the hormonal evaluation indicated a gradual decrease in E2, LH and P4 concentrations, starting from days 3, 7 and 10, respectively. No significant changes were noted in FSH or testosterone values. Besides, on day 10, prolactin concentrations remained unmodified. Even though most hormonal levels were found low on day 10, all values except E2 (undetectable) corresponded to the interval reported for the diestrus phase. Likewise, histological examination of ovarian tissue from nephrotic rats showed a considerable increase in the number of atretic follicles. These findings indicate that female rats with nephrotic syndrome develop an important endocrine dysfunction that probably involves steroidogenic tissues (ovary and/or adrenal gland), and suggest the existence of a hypothalamic-hypophyseal impairment.

Animals↗

Representation of tactile signals in primate supplementary motor area.

1. We have studied the neuronal activity in the supplementary motor area (SMA) of two monkeys who categorized the speed of moving tactile stimuli delivered to the glabrous skin of the hand ipsilateral to the site of cortical recording and contralateral to the responding arm. 2. A large number of SMA neurons responded to the stimuli of all speeds (176 of 522) but only when those stimuli controlled behavior. 3. A second class of SMA neurons responded differentially in the categorization task (35 during the stimuli and 51 during the reaction time period) and predicted its outcome. 4. To dissociate the interrupt target switches presses from the tactile categorization responses, sixteen neurons, which responded to the stimuli in all speeds, and 11 neurons, which discharged differentially, were tested in a visual control task. None of these two classes of neurons responded in this situation. 5. It is concluded that the SMA ipsilateral to sensory input and contralateral to the responding arm is involved in the sensory decision process in this somesthetic categorization task.

Afferent Pathways↗

Glycosaminoglycan-sulfate as plasma membrane component of pig spermatozoa.

The effect of specific glycosaminoglycan-hydrolyzing enzymes on the ruthenium red staining of pig spermatozoa was studied. Washed spermatozoa were incubated at 35 degrees C in buffer or with neuraminidase 0.5 units/ml, heparinase 0.2 mg/ml, or chondroitinase ABC 2.0 units/ml. After incubation sperm cells were washed, stained with ruthenium red and studied under the electron microscope. Anionic sites in the surface of untreated spermatozoa follow regularly the plasma membrane, but present are numerous processes constituting what has been defined as the glycocalyx. Neuraminidase did not affect the distribution of ruthenium red on the surface of the spermatozoa, but eliminated almost completely the processes of the glycocalyx. Heparinase caused loss of the ruthenium red-stained sites on the membrane surface of pig spermatozoa with less influence on the dense processes of the glycocalyx. A similar loss of ruthenium red-stained sites was observed with nitrous acid treatment. A striking effect of treatment with chondroitinase ABC was the production of a typical acrosome reaction.

Animals↗

Increased acrosome-reaction inducing activity of glycosaminoglycans by partial hydrolysis.

The possibility that partial hydrolysis of glycosaminoglycan-sulfates (GAGs) such as occurs during the last phases of follicular maturation could play some role in the activity of follicular fluid as an inducer of the acrosome reaction was explored. Hydrolysis of follicular fluid GAGs (ff-GAGs) for 30 min with low-pH HNO2 substantially increased (more than 3 times) its capacity to induce the acrosome reaction. This increase was significantly reduced when the time of hydrolysis was either shorter (10 min) or longer (60 min). Partial hydrolysis of spermatozoa GAGs by direct incubation of sperm cells with chondroitinase ABC was also capable of inducing the acrosome reaction.

Acrosome↗

Effect of divalent ions in acrosome reaction induced by glycosamineglycans in porcine spermatozoa.

Magnesium, calcium, and zinc at the concentration of 10 microM are capable of inducing a "true" acrosome reaction in the pig spermatozoa judged by the criteria of the fusion of the acrosome and the plasmatic membrane at the anterior region or the sperm nucleus. The optimal percent of acrosome reaction reached by any of the ions tested as a whole was 50%. When glycosamineglycan sulfate (GAGs) plus 10 microM of Mg++, Ca++, or Zn++ was added, they reach to 70-80% of acrosome reaction. At the electrom microscope, thin sections taken from pig spermatozoa treated with ions, GAGs, or ion + GAGs under optimal experimental conditions revealed the same pattern of acrosomal reaction. Results suggest the important role that divalent cations play in general in the induction of the acrosome reaction and question the so-called essential role of calcium ions.

Acrosome↗

Glycosamineglycan sulfate as acrosomal reaction-inducing factor of follicular fluid.

Follicular fluid from different mammalian species possesses two factors responsible for the induction of capacitation: a sperm-stimulating factor and an acrosomal reaction-inducing factor. The glycosamineglycan-sulfate (GAGs) extracted from pig follicular fluid induce acrosome reaction in pig spermatozoa which exhibit no morphological difference between the GAGs-induced reaction and the natural one. Acrosomal reaction commenced 30 min after the addition of GAGs and depended on GAGs concentration reaching 80% of acrosomal reacted spermatozoa after 6 hr of incubation with 7 mg of GAGs/ml. Chemical composition differs with the chemical data that characterize them as proteoglycans since those we obtained were practically protein free (2%). Another difference resides in the uronic acid content, which is almost twofold higher (59%). Electron microscope observations of the acrosomal reacted spermatozoa revealed that the addition of 10 mg/ml of trypsin soybean inhibitor did not interfere with any of the acrosomal reaction steps. The active capacitating factors may also originate from the follicular fluid released into the genital tract during ovulation.

Acrosome↗

Heparin-induced release of DNA template restrictions in human sperm zinc-depleted nuclei.

The addition of heparin to human sperm zinc-depleted nuclei releases DNA template restrictions. Spermatozoa depleted of zinc were assayed for (3H-methyl), thymidine incorporation was observed (27,500 +/- 1,248 dpm of 3H methyl-thymidine). Sperm cells incubated in the presence of 10 mg/ml of soybean trypsin inhibitor shows no effect in sperm nuclear swelling or in the release of DNA template restrictions. This process runs in a parallel fashion to the nuclear swelling induced by heparin, suggesting that swollen nuclei might be the source of DNA template. This was confirmed by autoradiographic studies, since all the sperm cells whose nuclei were judged swollen by morphological criteria also appeared labeled. The fact that there was no need for ATP generating system or of exogenous DNA polymerase emphasized the control role that zinc plays in the physiology of the human spermatozoa.

Cell Nucleus↗

Capacitation of human spermatozoa by heparin.

Zona-free hamster oocytes were incubated in vitro with human spermatozoa treated with heparin for 5, 10, 20, and 30 min. Sperm penetration was observed 2, 4, and 8 h after insemination. Sperm penetration was successful in 40%, 60%, and 69% of the oocytes treated for 10 min with heparin. Feminine and masculine pronuclei were clearly identifiable. In these samples, 80% of the sperm was alive and there was approximately 68% progress motility. Live sperm were observed for up to 24 h. Since glycosamineglycan sulfates are normally present in the ooplasm, zona pellucida, follicular and uterine fluids, this might suggest a direct participation of heparin in human sperm capacitation.

Animals↗

Heparin-induced nuclei decondensation of mammalian epididymal spermatozoa.

Decondensation of mammalian epididymal spermatozoa nuclei has been induced by exposure of intact spermatozoa to heparin, including those species in which ejaculated sperm were not susceptible to this treatment. This process occurred in the absence of any disulfide bond cleaving reactant. Swelling of caput epididymal spermatozoa nuclei commenced about 30 min after the addition of heparin, reaching 88% in rat, 33% in rabbit, 26% in pig, and 62% in bull of swelled nuclei after 6 hr of incubation at 37 degrees C with 5000 USP of heparin per ml. Corpus epididymal spermatozoa nuclei of rat and rabbit underwent decondensation at 50 degrees C reaching 24% and 22% of swelled nuclei, respectively, after 6 hr of incubation. The nuclei of the sperm cells of pig and bull from this epididymal region remained highly condensed as well as the nuclei of the cauda epididymal spermatozoa of all the species assayed. Electron microscope observations of the caput epididymal spermatozoa nuclei treated with heparin revealed that the chromatin is organized into nuclear bodies joined by a network of cross-linked and branched chromatin fibers in the species studied.

Animals↗

Decondensation of human sperm nuclei by glycosamineglycan-sulfate from sea urchin egg.

Decondensation of human and sea urchin spermatozoan nuclei has been induced by exposure of intact spermatozoa to glycosamineglycan-sulfate (GAGs) extracted from eggs of the sea urchin. Toxopneustes roseus. Swelling of human spermatozoan nuclei commenced about 60 min after the addition of GAGs and, depending on the extract concentration, the proportion of swollen nuclei reach 90% after 6 h of incubation when 20 mg of GAGs per ml was used. Swelling of sea urchin spermatozoan nuclei could be induced also with GAGs and/or with heparin. Swelling commenced about 10 min after the addition of GAGs/heparin reaching 90-92% after 60 min of incubation with 5 mg GAGs/500 USP heparin per ml. Preincubation of spermatozoa with 10 mg/ml of trypsin soybean inhibitor did not interfere with the swelling action of the GAGs.

Animals↗

Heparin binding sites in the human spermatozoa membrane.

The existence in the human spermatozoa membrane of receptorlike functional group for heparin was studied. Incubation of whole spermatozoa with tritiated heparin induced the specific binding of 745 +/- 112 pmol of heparin per 5 x 10(7) sperm cells with an intrinsic association constant KD = 3.6 x 10(-6) M. The specificity of binding was shown by the lack of competence in the binding process of some other glycosaminoglycans used at concentrations 20 x higher than heparin. However, dextran sulfate was a very efficient competitive agent. Autoradiography experiments showed that labeling was almost completely restricted to sperm cells in the process of nuclear decondensation. This technique showed the presence of a high amount of radioactive heparin in the isolated sperm membranes even after several washings. Heparin may participate both in the final part of the capacitation process (acrosome reaction) and in the decondensation of sperm nuclei.

Binding Sites↗

Differential distribution of cyclic-AMP receptors in human spermatozoa.

The binding properties of cyclic-AMP to intact and fractionated in heads and tails human spermatozoa have been studied. Whole spermatozoa bound 6.8 +/- 0.8 pmol of cyclic-AMP per 10(7) cells. Binding of cyclic-AMP was selectively located in the midpiece-tail region of the spermatozoa. Thus only 18% of the binding in the whole sperm could be accounted for by the binding on the sperm heads. This distribution is similar to that previously described for the protein kinase of bovine sperm. Blocking membrane sulfhydryl groups with pCMBS induced a drastic inhibition (65%) of the cyclic-AMP binding to the sperm heads, while only moderately reducing (30%) the binding in the sperm tails

4-Chloromercuribenzenesulfonate↗

Species specific decondensation of human spermatozoa nuclei by heparin.

Decondensation of human spermatozoa nuclei has been induced by exposure of intact spermatozoa to heparin, while the spermatozoa of rabbit, ram, and bull remained highly condensed under similar experimental conditions. This process occurred in the complete absence of any disulfide bond cleaving reactant. Swelling of human spermatozoa nuclei commenced about 30 min after the addition of heparin and depended on heparin concentration reaching 83% of swelled nuclei after 6 hr. of incubation with 5000 USP of heparin per ml. Addition of 10 mg/ml of trypsin soybean inhibitor did not interfere with the swelling action of heparin. Electron microscope observations of human spermatozoa nuclei treated with heparin revealed that the chromatin is organized into "hublike" nuclear bodies joined by a network of cross-linked and branched chromatin fibers ranging in thickness from 25 to 1.5 nm.

Animals↗