Biomedical subjects

R L Sprando

Publications and source records attributed to R L Sprando.

32 records · Page 2Linked to original sources

PubMed1990

Retinal cysts in the eye of the Asian musk shrew, Suncus murinus, (Mammalia, Insectivora).

The development of retinal cysts in the eye primordia of Suncus murinus embryos was studied. Retinal infoldings were first identified in the 17.5-day-old embryo (crown rump length--CRL = 6.3 mm). At this time, the apex of the developing retinal fold could be identified attached to the lens. The pigmented epithelium did not take part in the formation of the cyst. The retinal fold closed, to form a cyst, by day 23 of embryonic development (CRL = 14.0 mm). The newly formed retinal cyst remained continuous with the retina and extended toward the lens where it attached to the lens epithelium. Retinal cysts had an oval, bilaterally symmetrical shape and a narrow lumen, which, lined by the internal limiting membrane, occasionally contained cellular debris. The dimensions of the cysts were usually 100 to 150 microns (diameter) by 200 to 250 microns (length). Retinal cysts appeared in 45 per cent of the embryos examined.

Animals↗

PubMed1990

To what extent can spermatogenesis be maintained in the hypophysectomized adult rat testis with exogenously administered testosterone?

In a previous study it was demonstrated that spermatogenesis can be maintained quantitatively with exogenously administered testosterone in adult intact rats that lack LH. The studies described herein were designed to examine the extent to which spermatogenesis can be maintained quantitatively with exogenously administered testosterone in adult rats that lack all pituitary hormones. Adult male rats were hypophysectomized and testosterone was administered at the time of hypophysectomy via sustained release polydimethylsiloxane (PDS) capsules of increasing lengths. We used the PDS capsules to clamp testosterone at defined concentrations within the seminiferous tubule fluid over a 2- to 3-month treatment period. Mean testis weights and advanced spermatid numbers per testis stabilized by 8 weeks of testosterone treatment regardless of testosterone dose. Both testis weight and advanced spermatid number responded to testosterone dose, reaching plateaus of 1.2 g and 170 x 10(6) per testis, respectively. These values were 60% of, and significantly less than, the respective control values. This result was in striking contrast to the results of our previous study of LH-suppressed intact rats, in which exogenously administered testosterone resulted in testis weights and advanced spermatid numbers that plateaued at values not significantly different from those in controls. These different effects of testosterone in intact and hypophysectomized rats occurred despite the fact that the seminiferous tubule fluid testosterone concentrations achieved in the hypophysectomized rats (up to 25 ng/ml) were greater than the minimal testosterone concentration found previously to be required to maintain spermatogenesis quantitatively in LH-suppressed intact rats (13 ng/ml). Taken together, these results demonstrate clearly that intratesticular testosterone doses that are as high as or higher than those that maintain spermatogenesis quantitatively in intact rats lacking LH fail to maintain spermatogenesis quantitatively in rats lacking all pituitary hormones.

Animals↗

PubMed1990

Restoration of spermatogenesis by exogenously administered testosterone in rats made azoospermic by hypophysectomy or withdrawal of luteinizing hormone alone.

The major objective of the studies presented herein was to compare the extent to which exogenously administered testosterone is able to restore spermatogenesis in adult rats made azoospermic by withdrawal of all pituitary hormones (hypophysectomy for 4 weeks) vs. withdrawal of LH alone [testosterone- and estradiol-filled (TE) polydimethylsiloxane implants of 2.5 and 0.1 cm, respectively, for 8 weeks]. In hypophysectornized (Hypox) rats, serum LH and FSH were both undetectable; in the rats that received TE implants, serum LH was undetectable, but FSH was unaffected compared to control values. Seminiferous tubule fluid testosterone concentrations were reduced significantly from their control values of 60-65 to 1.4-1.7 ng/ml in the azoospermic Hypox and TE rats. These rats then received testosterone-filled implants of 4, 12, 18, or 24 cm and were killed 2 months later. In both the Hypox and TE rats, seminiferous tubule fluid testosterone concentrations rose linearly with increasing capsule sizes, and with each of the implant sizes, these concentrations did not differ significantly between the Hypox and TE rats. This made it possible for the first time to examine the effects of comparable intratesticular testosterone concentrations on the numbers of advanced spermatids per testis that could be restored in the azoospermic testes of rats lacking all pituitary factors vs. those lacking only LH. The results that we present demonstrate that the numbers of restored advanced spermatids were consistently and significantly lower in Hypox than in TE rats despite equivalent seminiferous tubule fluid testosterone concentrations. These results provide quantitative conclusive evidence to support the contention that pituitary factors in addition to LH are required for the quantitative restoration of spermatogenesis in the adult rat testis.

Animals↗

PubMed1990

Reversal of long-term LH deprivation on testosterone secretion and Leydig cell volume, number and proliferation in adult rats.

The purpose of this study was to determine whether Leydig cell volume and function could recover fully from long-term LH deprivation upon restoration of endogenous LH secretion, and whether the restoration of LH would elicit a mitogenic response, i.e. stimulate Leydig cell proliferation or affect Leydig cell number per testis. LH secretion was inhibited by treating adult rats with testosterone and oestradiol-filled (TO) silicone elastomer implants (16 weeks), and was restored by removing the implants. Changes in serum concentrations of LH and FSH, LH-stimulated testosterone secretion by testes perfused in vitro, Leydig cell volume and number per testis, average Leydig cell volume and Leydig cell [3H]thymidine incorporation were measured at weekly intervals following implant removal. The TO implants inhibited (P less than 0.01) LH secretion, but serum concentrations of FSH were not significantly different (P greater than 0.10) from control values. After implant removal, serum LH returned to control values within 1 week, whereas serum FSH increased twofold (P less than 0.01) and returned to control values at 4 weeks. LH-stimulated in-vitro testosterone secretion was inhibited by more than 99% in TO-implanted rats, but increased (P less than 0.01) to 80% of control values by 8 weeks after implant removal. The total volume of Leydig cells per testis and the volume of an average Leydig cell were 14 and 19% of control values respectively, after 16 weeks of TO implantation (P less than 0.01), but returned to 83 and 86% of controls (P greater than 0.10) respectively, by 6 weeks after implant removal. Leydig cell proliferation ([3H]thymidine labelling index) was low (less than 0.1%) in both control and TO-implanted rats, increased (P less than 0.01) fivefold from 1 to 4 weeks after implant removal and then declined to control values at 6 weeks. The increase in Leydig cell [3H]thymidine incorporation was mimicked by treating TO-implanted rats with exogenous LH, but not FSH. Leydig cells were identified in both the interstitium and the lamina propria of the seminiferous epithelium. The proportion of Leydig cell nuclei in the lamina propria was 30-fold greater (P less than 0.01) at 1 and 3 weeks after implant removal (3%) compared with that for control and TO-implanted rats (0.1%). Total Leydig cell number per testis was marginally but not significantly (P = 0.06) decreased in rats treated with TO implants for 16 weeks when compared with controls (18.4 +/- 2.2 vs 25.4 +/- 1.2 x 10(6)).(ABSTRACT TRUNCATED AT 400 WORDS)

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PubMed1989

Restoration of advanced spermatogenic cells in the experimentally regressed rat testis: quantitative relationship to testosterone concentration within the testis.

We examined the effect of exogenously administered testosterone (T) on the quantitative restoration of advanced spermatogenic cells in adult rat testes rendered azoospermic by treating rats with polydimethylsiloxane (PDS) implants of T and estradiol (E). Experimental rats received PDS-TE implants for an initial 8-week period; control rats received empty implants. By 8 weeks of PDS-TE treatment, rats became severely oligospermic, and the T concentration within the seminiferous tubule fluid (STF) was reduced approximately 80% (from 57.8 ng/ml in controls to 9.6 ng/ml). After the initial 8-week PDS-TE treatment, PDS-TE implants were removed from one group of rats; a second group of PDS-TE-implanted rats received an additional PDS-T implant of 24 cm. Eight weeks after the removal of PDS-TE implants or the implantation of additional T, testis weight and numbers of advanced spermatogenic cells were restored to those of control rats. The STF T concentration 8 weeks after the removal of PDS-TE implants also was restored to that in control rats. In contrast, the STF T concentration increased to only 40% of control values in the rats that received an additional T implant. Despite this 60% reduction in T concentration compared to the control value, advanced spermatogenic cell number was restored to a value indistinguishable from that of intact controls. These observations indicate that spermatogenesis can be quantitatively restored in PDS-TE-implanted rats with exogenously administered T, and moreover, that this restoration does not require the high T concentration found in the STF of intact control rats.

Animals↗

PubMed1988

Spermiogenesis in the red-ear turtle (Pseudemys scripta) and the domestic fowl (Gallus domesticus): a study of cytoplasmic events including cell volume changes and cytoplasmic elimination.

Nuclear and cytoplasmic volume changes as well as the elimination of residual spermatid cytoplasm were investigated in the red-ear turtle (Pseudemys scripta) and the rooster (Gallus domesticus). Nuclei of newly formed spermatids which were originally centrally located became eccentrically located within the cell in both species. Shortly thereafter the nuclear pole of the spermatid was found situated within deep crypts of a Sertoli cell. The cytoplasm of elongating spermatids was displaced along the nonacrosomal region of the nucleus and the proximal flagellum. In both species sheetlike Sertoli cell processes indented spermatid cytoplasm adjacent to the nucleus and appeared to segregate small packets of the cytoplasm. In the turtle, these packets of cytoplasm were separated from the spermatid. In both the turtle and rooster, a portion of the spermatid cytoplasm was displaced forward over the acrosomal region of the spermatid to resemble a hood. As spermatids were transported to the seminiferous tubular lumen, cytoplasmic lobes which projected forward of the spermatid head were formed by preferential flow of cytoplasm into one aspect of the cytoplasmic hood. In both species, at sperm release the cytoplasmic lobe was disengaged from the spermatid head to form a large residual body that was internalized and degraded within the Sertoli cell. Medium-sized cytoplasmic lobes were pinched from the head and neck region of the turtle and rooster spermatids, respectively. In the turtle, small-sized mitochondrial-rich cytoplasmic fragments budded from the caudal head and midpiece of the spermatids and were phagocytosed by the Sertoli cell. Thus, cytoplasmic elimination occurred through 1) segregation of cytoplasmic packets by Sertoli penetrating processes (turtle), 2) elimination of large and medium-sized residual bodies from the head (turtle and bird), and 3) budding of small mitochondrial-rich cytoplasmic fragments from the region of the midpiece (turtle). In the turtle a 79% reduction in total cell volume occurred during spermiogenesis which was the result of an 84% cytoplasmic reduction and a 78% nuclear reduction. During spermiogenesis, the rooster lost 97% of its total cell volume due to a 97% cytoplasmic volume change and a 96% nuclear volume change.

Animals↗

PubMed1988

Spermiogenesis in the bluegill (Lepomis macrochirus): a study of cytoplasmic events including cell volume changes and cytoplasmic elimination.

The process involved in the reduction of both nuclear and cytoplasmic volume was investigated in the bluegill (Lepomis macrochirus), a teleost fish. Young spermatids contained centrally positioned nuclei which, with time, moved toward the cell surface to become eccentrically positioned. Chromatin condensation was initiated from a region near the implantation fossa, whereas at the opposite pole of the nucleus an area sparse in heterochromatin (clear area) was noted. The nuclear membrane lying adjacent to the clear area dissolved and subsequently reformed, yielding a nucleus with a reduced volume. During this process, packets of cytoplasm surrounded by a double membrane were formed along the future midpiece. The packets of cytoplasm migrated toward the cell surface, protruded from the surface, and were extruded into the spermatocyst lumen. These structures, termed residual bodies, were subsequently endocytosed, accumulated into large phagocytic vacuoles, and eventually degraded by the nearby Sertoli cell. When the spermatocyst ruptured, spermatozoa containing sparse cytoplasm were released into the excurrent duct system. During spermiogenesis, both the nuclear and cytoplasmic volumes decreased substantially (80%, 92% respectively) leading to an overall 87% reduction in total cell volume.

Animals↗

PubMed1988

Spermiogenesis in the bullfrog (Rana catesbeiana): a study of cytoplasmic events including cell volume changes and cytoplasmic elimination.

The process by which spermatid cytoplasmic volume is reduced and cytoplasm eliminated during spermiogenesis was investigated in the bullfrog Rana catesbeiana. At early phases of spermiogenesis, newly formed, rounded spermatids were found within spermatocysts. As acrosomal development, nuclear elongation, and chromatin condensation occurred, spermatid nuclei became eccentric within the cell. A cytoplasmic lobe formed from the caudal spermatid head and flagellum and extended toward the seminiferous tubule lumen. The cytoplasmic lobe underwent progressive condensation whereby most of its cytoplasm became extremely electron dense and contrasted sharply with numerous electron-translucent vesicles contained therein. At the completion of spermiogenesis, many spermatids with their highly condensed cytoplasm still attached were released from their Sertoli cell into the lumen of the seminiferous tubule. There was no evidence of the phagocytosis of residual bodies by Sertoli cells. Because spermatozoa are normally retained in the testis in winter and are not released until the following breeding season, sperm were induced to traverse the duct system with a single injection of hCG. Some spermatids remained attached to their cytoplasm during the sojourn through the testicular and kidney ducts; however, by the time the sperm reached the Wolffian duct, separation had occurred. The discarded cytoplasmic lobe (residual body) appeared to be degraded with the epithelium of the Wolffian duct. It was determined that the volume of the spermatid was reduced by 87% during spermiogenesis through a nuclear volume decrease of 76% and cytoplasmic volume decrease of 95.3%.

Animals↗

PubMed1987

Comparative study of cytoplasmic elimination in spermatids of selected mammalian species.

In the transformation of a spermatid into a spermatozoon, cytoplasmic loss occurs, allowing the sperm to swim unhindered. Cytoplasmic loss takes place via elimination of the residual body and through structures known as tubulobulbar complexes. A determination of cytoplasmic loss in several species was undertaken by using high-resolution electron microscopic morphometric techniques. During the period that tubulobulbar complexes are present, an average 53% cytoplasmic loss was recorded for five species (guinea pig, 48.8 +/- 6.2% (SEM); monkey, 60.3 +/- 4.3%; opossum, 54.5 +/- 4.4%; rabbit, 46.9 +/- 2.7%; and rat, 55.7 +/- 4.9%), whereas there was essentially no loss or gain in cytoplasm during the same period for round spermatids. Surprisingly, during spermatid elongation an approximate 36% loss of cytoplasm was also recorded for five species (guinea pig, 50.1 +/- 6.3%; monkey, 30.0 +/- 15.4%; opossum, 25.4 +/- 9.0%; rabbit, 42.4 +/- 8.6%; and rat, 34.9 +/- 11.9%), which is only partially (approximately 60%) accounted for by fluid pumping from the nucleus during nuclear condensation. A densification of the cytoplasm of elongate spermatids, as compared with round spermatids, suggests fluid is also pumped from the elongating spermatid cytoplasm. Fluid loss from germ cells may contribute to the seminiferous tubule fluid, a fluid previously thought to be solely of Sertoli-cell origin.

Animals↗

PubMed1987

A comparative study of Sertoli cell ectoplasmic specializations in selected non-mammalian vertebrates.

Ectoplasmic specializations (ES) facing spermatids were studied in species representative of four classes of non-mammalian vertebrates (Pisces--bluegill; Amphibia--bullfrog; Reptilia--red eared turtle; Aves--domestic chicken). ES was not seen in the bluegill but was present in all other species studied. In the frog, turtle, and chicken, ES did not resemble its mammalian counterpart and could only be characterized by the presence of 6 nm filaments (presumedly actin) within the somatic cell facing the head region of elongating spermatids. ES filaments were sparse in the frog and were sometimes associated with more deeply situated endoplasmic reticulum. Turtle ES filaments were abundant and encircled the acrosomal region of the spermatid head and were delimited by fenestrated saccules of endoplasmic reticulum. In the chicken, ES filaments were prominent but less abundant than in the turtle. Six nanometer filaments of the chicken ES appeared in a tangled mass and were not associated with clearly defined endoplasmic reticulum. In the three species where ES was found, it first developed as spermatids became entrenched within the surrounding somatic cell. Neither cell elongation, nuclear elongation, or movement of the nucleus to the cell surface was synchronized with the onset of ES development. That ES development was seen concomitant with spermatid entrenchment and spermatid orientation suggested a role for ES in these processes. This hypothesis was further strengthened by observations in the fish where ES was lacking and where spermatid entrenchment within the somatic cell, did not occur. The study also supported the hypothesis that ES acts as a cytoskeletal mantle to which other cytoskeletal elements within the cell interact to affect the position of elongate spermatids within the epithelium. The dissolution of ES prior to spermiation and concomitant loss of a close relationship between cells suggests that ES is also related to somatic cell-germ cell adhesion and therefore plays an important role in the spermiation process.

Animals↗

PubMed1987

Germ cell-somatic cell relationships: a comparative study of intercellular junctions during spermatogenesis in selected non-mammalian vertebrates.

Specialized germ cell-somatic cell relationships were surveyed in the testis of species representative of four classes of non-mammalian vertebrates. Desmosome-like junctions were present in all classes studied. In the teleost fish studied (bluegill; Lepomis macrochirus), small, infrequent desmosomes, seen between the spherical cyst cells and spermatocytes, were characterized by poorly represented subsurface densities. In the bullfrog (Rana catesbeiana), similar desmosome-like junctions were found between cyst cell processes and spermatocytes. Reptilian (turtle; Pseudameys scripta) desmosome-like junctions between Sertoli cells and germ cells were heterogeneous and more numerous than those junctions found in fish and amphibians. In general, the reptilian desmosome-like junctions were extensive structures displaying 10 nm filaments associated with the Sertoli cell component of the junctions. Regions within the desmosome where the two plasma membranes converged suggested that gap junctions were a component of the desmosome-like junctions. "Desmosome-gap" junctions persisted in turtle spermatids for sometime after nuclear elongation had commenced. In birds (chicken; Gallus domesticus), "desmosome-gap" junctions, similar to those seen in turtles were described between both spermatocytes and Sertoli cells, and spermatids and Sertoli cells. These junctions were frequently lined by saccules of endoplasmic reticulum. The presence of gap junctions suggest the evolution of mechanisms for somatic cell-germ cell communication although more species should be examined to confirm this hypothesis.

Animals↗

PubMedDate not supplied

Developmental toxicity of orange B when given to rats by gavage.

The pyrazolone dye Orange B was given by gavage to pregnant Osborne-Mendel rats throughout gestation. Dose levels of 0, 15, 30, 100, 200, 400, or 700 mg/kg body weight were given daily. On gestation day 20, the females were killed and cesarean sections were performed. Feed consumption and maternal weight gain were not affected. No dose-related changes were seen in maternal clinical findings, implantations, fetal viability, or fetal size (weight and length). No compound-related effects were seen in sternebral development. No dose-related effect was seen in the incidence of skeletal variations in fetuses or in the number of litters containing fetuses with skeletal variations. Skeletal development, as measured by the average number of ossified vertebrae, was similar in all groups. No compound-related effects were seen in soft-tissue development.

Administration, Oral↗

PubMedDate not supplied

Does ethane 1,2-dimethanesulphonate (EDS) have a direct cytotoxic effect on the seminiferous epithelium of the rat testis?

The authors examined the possibility that ethane 1,2-dimethanesulphonate (EDS) has a cytotoxic effect on spermatogenesis that is not secondary to androgen withdrawal resulting from the well known cytotoxic effect of EDS on Leydig cells. Adult male rats were implanted with polydimethylsiloxane (PDS) capsules containing testosterone (T) and estradiol (E), and were simultaneously injected with EDS. The PDS-TE implants, by inhibiting luteinizing hormone (LH) production, prevented Leydig cells from repopulating the testis and clamped testosterone within the seminiferous tubules at increasing concentrations relative to implant size. In rats that received EDS alone, the number of advanced spermatids per testis was significantly reduced by 2 weeks, but within 8 weeks returned to the numbers maintained in vehicle-injected control rats or in vehicle-injected rats that received testosterone- and estradiol-filled capsules of 24 cm and 0.1 cm, respectively (PDS-24TE). Surprisingly, in rats that received an EDS injection plus PDS-24TE implants, the number of advanced spermatids per testis was significantly reduced at 8 weeks and severe seminiferous tubule atrophy occurred despite the fact that the testosterone concentration was sufficient to quantitatively maintain spermatogenesis in vehicle-injected rats. In rats injected with EDS and implanted with 24 cm testosterone but not estradiol-filled capsules (PDS-24T), the advanced spermatid number per testis was significantly higher than that in the EDS plus PDS-24TE rats, but significantly lower than that in control rats. These results suggest that EDS may have a cytotoxic effect on the seminiferous epithelium that is independent of the elimination of Leydig cells, and the EDS and estradiol act synergistically to exert a profound toxic effect on spermatogenesis.

Animals↗

PubMedDate not supplied

Is Leydig cell steroidogenic function affected by the germ cell content of the seminiferous tubules?

The effect of testicular germ cell content on Leydig cell steroidogenic function in vivo in adult rats was examined. Three experimental paradigms were used to effect germ cell changes. First, a vitamin A-depletion/repletion regimen was used to achieve synchrony at different stages of the cycle of the seminiferous epithelium and thus produce testes with widely differing germ cell contents. Second, long term vitamin A depletion was used to effect germ cell, but not Leydig cell, loss. Third, Leydig cells and germ cells first were eliminated from the testes of adult rats by the administration of ethane 1,2-dimethane sulfonate (EDS) along with testosterone- and estradiol-filled Silastic capsules; Leydig cells were then restored to the germ cell-depleted testes by removal of the luteiniging hormone (LH)-suppressing capsules. Serum, interstitial fluid, and seminiferous tubule fluid testosterone concentrations did not differ between rats in which at least 70% of the seminiferous tubules contained germ cells at stages VII-VIII or at stages XII-III of the cycle. The capacity of the testes of these rats to produce testosterone, assessed by their in vitro perfusion with maximally stimulating LH, also showed no differences despite the differences in germ cell content. Elimination of germ cells throughout the testes by long term vitamin A depletion also did not affect the steroidogenic function of the testes. Finally, the steroidogenic function of Leydig cells restored to germ cell-depleted testes was indistinguishable from that of germ cell-containing controls. These results, taken together, provide evidence supporting the contention that the germ cell content of the testis has little or no effect on testicular steroidogenic function.

Animals↗