PubMed Health⌕ Search

Biomedical subjects

M C Poole

Publications and source records attributed to M C Poole.

14 recordsLinked to original sources

The effects of space flight during gestation on rat uterine smooth muscle.

Pregnant rats were flown on a NASA space shuttle during days 11-20 of gestation and the effects of pregnancy maintenance and uterine myometrial smooth muscle were evaluated. There was no effect of space flight on fetal mass in rats examined at recovery on day 20 of gestation (G-20). Rats allowed to go to term delivered vaginally at days 22-23, but space flight significantly decreased pup mass at birth. Space flight did not alter myometrial smooth muscle volume at G-20 and postpartum when compared to flight delayed synchronous controls, but flight animals showed a 37% decrease in myometrial smooth muscle volume between G-20 and postpartum. In contrast, smooth muscle volume of the myometrium of controls, postpartum, was similar to G-20. We conclude that myometrial smooth muscle, which hypertrophies during space flight similar to controls, reacclimates acutely to earth's gravity between G-20 and parturition with a dramatic reduction in volume.

Animals↗

Estrogen affects performance of ovariectomized rats in a two-choice water-escape working memory task.

To determine if estrogen would protect treated rats from deficits in performance on a working memory task across time, 18 female 6-month-old Sprague-Dawley rats were trained to a criterion on a water-escape spatial delayed matching-to-sample problem. Following training, rats were ovariectomized, and nine were maintained on estrogen (polyestradiol-phosphate, 0.5 mg every 3 weeks) and nine on its vehicle for 200 days. After recovery from surgery, the rats were tested for performance every 6 weeks under three conditions: 5 min retention interval (RI); 30 min RI; and 30 min RI with an emotional experience during the RI. Analysis of correct choices revealed that estrogen-treated rats made more correct choices (p < .05) than controls on the 5 min undisturbed interval; estrogen tended to impair performance on the emotionally distracting interval. Estrogen apparently protected working memory on the undisturbed trials and might be pertinent to the maintenance of memory in female mammals.

Animals↗

An image processing/stereological analysis system for transmission electron microscopy.

This study examines the feasibility of combining computer image digitization, image enhancement, and point counting stereological techniques to quantify video images from transmission electron microscopes (TEM). The essential hardware consists of an IBM PC/AT, a Matrox imaging board, a digitizing tablet, a high resolution black and white monitor, and a portable mass storage device. In addition a video camera must be mounted to the TEM. The software is written in three modules which have numerous routines for image acquisition, enhancement, and quantification. Quantification is achieved by selecting an electronic lattice and superimposing it on the cell image. A cursor is moved on the lattice (via the digitizing tablet) and the points are entered into a spreadsheet. One of the major limitations of the system was the reduced resolution inherent in the current hardware. However, sampling experiments showed that one could compensate for the reduced resolution by increasing the magnification of the digitized images, and the stereological values from digitized images compared favorably to those from electron micrographs. Furthermore, the system proved advantageous by eliminating the usual darkroom work, and in enhancing low contrast tissue. In spite of several hardware limitations, the concept of quantifying computer digitized TEM images appears promising.

Animals↗

Alteration of the mammotroph Golgi complex by the dopamine agonist 2 Br-alpha-ergocryptine (CB-154) in ovariectomized estrogen primed rats.

The present study examined the acute effects of 2 Br-alpha-ergocryptine (CB-154, a dopamine agonist) on mammotroph organelles during prolactin (PRL) suppression. Ovariectomized estrogen-primed rats received a single injection (sc) of 0.5 mg CB-154 and the animals were killed at intervals following injection. The anterior pituitary glands were fixed for electron microscopy and immunocytochemistry was used to confirm mammotroph identification. Serum PRL levels were determined by RIA. Following CB-154 administration, serum PRL was significantly (P less than 0.05) reduced within 15 minutes and was suppressed (P less than 0.01) to ovariectomized levels at 2 and 6 hours. A stereological analysis of mammotrophs in the central regions of the anterior pituitary showed that the Golgi complex volume was significantly (P less than 0.05) reduced at 2 hours after CB-154 treatment. However, the Golgi complex volume had recovered by 6 hours post CB-154 injection. In addition, the volumes of the mammotroph cells, the mature secretory granules, and the secondary lysosomes had significantly increased by 6 hours. There were no significant changes in any of the organelles following CB-154 in the mammotrophs from the peripheral regions of the gland. These studies show that the Golgi complex is especially susceptible to acute morphological changes induced by bromocryptine and that the mammotrophs in the central regions are more responsive to CB-154 than those in the peripheral regions.

Animals↗

Glucose affects in vitro maturation of fetal rat islets.

Fetal pancreatic islets (21.5 days old) were cultured in RPMI 1640 containing either 2.8 or 11.1 mM glucose for 7 days. After the 7-day culture period, islets cultured in 2.8 mM glucose demonstrated a minimal first phase of insulin secretion in response to acute glucose stimulation, whereas islets cultured in 11.1 mM glucose demonstrated a biphasic insulin secretory pattern. Islets cultured in 11.1 mM glucose initiated insulin secretion at 4.4 +/- 0.1 mM glucose and plateaued at 11.6 mM glucose when exposed to a linear gradient. In addition, culture in 11.1 mM glucose increased DNA content (P less than 0.01) and [3H]thymidine incorporation (P less than 0.05) in fetal islets. However, ultrastructural morphometric analysis indicated that the actual number of beta-cells within islets cultured in either 2.8 or 11.1 mM glucose did not increase. The insulin contents of islets cultured in 2.8 and 11.1 mM glucose were 0.46 +/- 0.06 and 1.14 +/- 0.10 mU/islet, respectively. During subsequent glucose stimulation, islets cultured in 2.8 and 11.1 mM glucose released 3% and 5.6% of their total insulin content, respectively. Ultrastructural morphometric analysis indicated that 11.1 mM glucose stimulated an increase in the volume of individual beta-cells, i.e. hypertrophy. The hypertrophy of beta-cells within islets cultured in 11.1 mM glucose resulted in a concomitant increase in islet volume. Finally, the hypertrophy of beta-cells within islets cultured in 11.1 mM glucose was a result of increased volumes of mitochondria, secretory granules, and, to the greatest extent, endoplasmic reticulum. These findings indicate that glucose is a potent factor in the maturation of cultured fetal rat islets.

Animals↗

Effects of growth hormone on the in vitro maturation of fetal islets.

To study the effects of growth hormone (GH) on the in vitro maturation of fetal islets, the fetal islets were cultured for 7 days in RPMI 1640 containing 10% fetal bovine serum and 11.1 mM glucose with or without GH. Culture with 1 microgram/ml of bovine GH increased the DNA content of the islets and [3H]thymidine incorporation into DNA confirming results of other investigators. In addition, however, the insulin secretory dynamics and ultrastructural morphometrics were investigated. It was found that GH-treated islets demonstrated increased insulin release during acute glucose stimulation when expressed as microunits per islet per minute. However, when insulin release during acute glucose stimulation was expressed as microunits per microgram of DNA per minute to compensate for the increased DNA content of GH-treated islets, no change in insulin release was observed compared to control islets. When GH-treated islets were perifused with a linear glucose gradient, the insulin secretory response was suppressed as indicated by changes in the threshold level, plateau level, and half-maximal response. Ultrastructural morphometric data showed that the average beta-cell volume in control and GH-treated islets was the same, eliminating the possibility that beta-cell hypertrophy occurred. Similarly, the nuclear volumes of the beta cells in control and GH-treated islets remained unchanged. This finding coupled with the observed increased DNA content and [3H]thymidine incorporation suggests that GH functions by increasing cell multiplication within the islets and not by inducing polyploidy. Finally, the volumes of cytoplasmic organelles in control and GH-treated islets were the same indicating that cytodifferentiation did not occur.

Animals↗

A computer program using a nested analysis of variance in morphometric sampling.

The quality of the sample set is absolutely essential in a morphometric analysis. In this paper a computer program is described which uses a nested analysis of variance in defining the sample. The program denotes those levels of the sampling protocol at which additional sampling should be performed to reduce the variance within the sample set. Furthermore, the program is written for use on a microcomputer which allows it to be used routinely in monitoring a sample set.

Analysis of Variance↗

Cellular distribution within the rat adenohypophysis: a morphometric study.

A morphometric analysis of the cell types within the adenohypophysis of the female rat was conducted with an emphasis on regional variations. Type I and type II gonadotropes and corticotropes had the largest volumes, mammotropes and somatotropes were of medium size, and thyrotropes and chromophobes were the smallest cells. There was no regional variation in the volumes of the respective cell types. Somatotropes and mammotropes were most numerous, followed by the gonadotropes, chromophobes, corticotropes, and thyrotropes. There was no significant regional localization of the somatotropes, mammotropes, corticotropes, and thyrotropes. Gonadotropes were best defined as two populations with type I cells being localized in the central regions and type II cells being distributed throughout the gland. Chromophobes were the only cells with a significant distribution in the anterior regions and were most numerous in the anterior peripheral regions. These fine-structural morphometric findings are discussed in relation to other studies of the adenohypophysis that utilize immunocytochemistry.

Animals↗

Morphometric analysis of the autophagic and crinophagic lysosomal systems in mammotropes throughout the estrous cycle of the rat.

The crinophagic and autophagic lysosomal systems were studied in mammotropes (prolactin secreting cells) of the adenohypophysis throughout the estrous cycle of the rat. By means of morphometric analysis, it was found that the volume of secondary autophagic lysosomes was usually greater than that of the crinophagic type. Although the volumes of both secondary autophagic and crinophagic lysosomes were minimal throughout proestrus and diestrus 2, the autophagic lysosomal volume per mammotrope was elevated during the estrous period. The volume of secondary crinophagic lysosomes per mammotrope increased during late estrus and remained elevated throughout early diestrus 1. Furthermore, there was an inverse relationship between the volume of mature secretory granules per cell and of the crinophagic system. These data suggest a role for lysosomes in the regulation of synthesis and secretion of prolactin by the adenohypophysis of the rat.

Animals↗

Intracellular dynamics in pituitary mammotropes throughout the rat estrous cycle. I. Morphometric methodology and hormonal correlations with cellular and nuclear volumes.

Serum prolactin (PRL) levels display regular patterns throughout the 4-day rat estrous cycle. A morphometric analysis of mammotropes (PRL-sereting cells) in the anterior pituitary was performed in an effort to quantify some of the ultrastructural events occurring prior to and during the secretion of PRL throughout the estrous cycle. In this study, a systematic sampling protocol applicable to the rat adenohypophysis was developed, which took into account the lack of synchrony in estrous cycles among the animals, the heterogeneity of the gland, and the number of cells required to yield a representative sample. These techniques were used to determine the cell, nuclear, nucleolar, and cytosol volumes, which were correlated with the changing levels of serum estradiol and PRL and pituitary PRL. Estradiol concentrations rose during late diestrus-1 and continued rising in a pulsatile manner, reaching maximal levels during proestrus. The elevations of estrogen were accompanied by increases in nucleolar and nuclear volumes. However, at early estrus and on the afternoon of diestrus-2, times at which it has been reported that progesterone levels are elevated, there was a dampening effect and these volumes declined. During early diestrus-2, when estradiol levels began to rise, there were increases in nucleolar, nuclear, cellular and cytosol volumes, as well as increases in pituitary PRL; but at late diestrus-2 there was a marked increase in nuclear and nucleolar volumes which was not accompanied by increases in cellular and cytosol volumes. During proestrus, two peaks of serum PRL were found, at 6 pm and 12 m, which were concomitant with increased nucleolar, nuclear and cellular volumes. The 6-pm proestrous peak of PRL was preceded by elevated pituitary PRL; however, the 12-m proestrous peak occurred when the pituitary PRL was minimal, even though nuclear and cellular volumes were elevated. This suggests that the 6-pm release of PRL was derived from previously synthesized and stored hormone, while the 12-m peak was the result of the immediate release of PRL, with little storage.

Animals↗

Intracellular dynamics in pituitary mammotropes throughout the rat estrous cycle. II. Changes in synthetic and secretory organelles.

The organellar dynamics within mammotropes (prolactin secreting cells) of the anterior pituitary were examined using morphometry throughout the four-day rat estrous cycle. Two predominant periods of maximal organellar activity were found. The first period, which occurred during proestrus, resulted in two peaks of serum PRL at 6 pm and 12 m proestrus, while the second occurred during middiestrus-2 and resulted in an increase of pituitary PRL, but not a major serum PRL peak. The mitochondrial volume rose prior to that of other organelles during early proestrus, rising 110% from 4 am late diestrus-2 to 6 pm proestrus. However, the number of mitochondria per cell remained constant throughout the cycle. The volumes of rough endoplasmic reticulum (RER) and Golgi complex reached maximal proportions at 8 pm and 12 m proestrus, respectively. The RER volume was 153% greater than at 4 am late diestrus-2, while the Golgi complex had increased by 200%. We suggest that changes in the RER and Golgi complex volumes were the result of two processes: the synthesis of new membrane and the swelling of existing complexes. This study further substantiated that the serum peak of PRL at 6 pm proestrus is the result of stored mature granules being released, whereas the PRL peak at 12 m proestrus is due to the immediate release of hormone without any storage.

Animals↗

A computer program for the morphometric analysis of cell profiles.

A computer program which yields values for the volumes, surface areas, and volume/surface area ratios of cell profiles is described for use on a desktop calculator (minicomputer). This program uses standard morphometric procedures, and incorporates data obtained from electron micrographs at two levels of sampling. The main program yields values for the 'average cell volume' at the tissue level of sampling. Two options at the cellular level of sampling are also included which yield values for the volumes, surface areas and volume/surface area ratios for the organelles. The first option allows an analysis of 'whole cells' containing equatorial profiles through the nucleus, while the second option permits a 'fractional' approach using segments of the cells. Finally, some of the advantages of the two options are discussed.

Cell Count↗