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PubMed2002

Tyrosine hydroxylase-immunoreactive interneurons in the olfactory bulb of the frogs Rana pipiens and Xenopus laevis.

We studied tyrosine hydroxylase (TH)-immunoreactive neurons and neuropil in the olfactory bulb of the leopard frog, Rana pipiens, and in the clawed frog, Xenopus laevis. In both frogs, TH processes in the main olfactory bulb showed a trilaminar organization, with a densely stained external glomerular layer (GL), a moderately stained middle mitral cell layer (MCL), and internally a weakly stained internal plexiform layer (IPL) and granule cell layer (GRL). TH-positive cells in the MCL and IPL could be divided into two types. Type 1 cells had one or two thick dendrites that arborized within glomeruli in the GL and often had a thin "axon-like" process that exited the cell on the internal surface, with a recurrent collateral that ascended into the GL. Type 2 cells had beaded dendrites arborizing in the MCL and no discernible axons. Both type 1 and type 2 cells were numerous in the MCL and IPL of Rana, whereas only type 2 cells were common in the MCL and IPL of Xenopus. In the GL, labeled cells were numerous in Xenopus but rare in Rana. Mitral cells were stained retrogradely by tracer injection into the lateral olfactory tract and by local injection into the bulb. In no case was double labeling for TH observed, suggesting that TH-positive cells in frog olfactory bulb are likely to be interneurons. Double labeling with an anti-gamma-aminobutyric acid (GABA) antibody showed that the TH-positive cells formed a population separate from the GABA-containing interneurons.

Animals↗
PubMed1981

The aberrant retino-retinal projection during optic nerve regeneration in the frog. I. Time course of formation and cells of origin.

We have reported previously that during optic nerve regeneration in Rana pipiens, axons are misrouted into the opposite nerve and retina. In the present investigation we have examined the time course of formation of these "misrouted" axons and their cells of origin. The right eye of 31 frogs was injected with 3H-proline at various times after right optic nerve crush. In every frog examined 2 weeks and later after nerve crush, the distribution of autoradiographic label indicated that axons from the right eye had grown into the left optic nerve at the chiasm. The amount of label increased from 2 weeks to reach a maximum at 6 weeks where the entire left nerve was filled with silver grains. At 5 to 6 weeks after crush, labeled axons were found within the ganglion cell fiber layer (GCFL) of the retina of the opposite eye for a maximum distance of 2.3 mm from the optic disc. In frogs examined at intervals later than 6 weeks after crush, the amount of label within the left eye and nerve progressively decreased, indicating a gradual disappearance of the misrouted axons. Studies using anterograde transport of horseradish peroxidase (HRP) after nerve injection confirmed these autoradiographic findings. The position of ganglion cells in the right eye whose axons were misrouted to the left eye was determined by retrograde transport of HRP. Five or 6 weeks after crushing the right optic nerve, the left eye was injected with HRP and labeled ganglion cells were found throughout the right eye retina. The largest percentage of labeled cells was found within the ventral half of the retina, particularly within the temporal quadrant, and nearly all of the labeled cells were found in more peripheral portions of the retina. Since few retino-retinal axons are found during normal development, the present results show that the factors guiding regenerating axons in the adult frog differ substantially from those present during development.

Animals↗
PubMed1987

Evidence for parallel processing in the frog's auditory thalamus.

We have conducted anatomical and physiological experiments to investigate the functional organization of the dorsal thalamus in the northern leopard frog (Rana pipiens pipiens). Our studies provide evidence for parallel auditory processing at this level of the frog's brain. Acoustically evoked potentials were recorded from the posterior and central thalamic nuclei and several differences in sound-evoked activity were noted between them: the amplitude of acoustically evoked potentials (AEPs), in response to a standard search stimulus, was always greater in the central, as opposed to the posterior, nucleus; the posterior, but not central, nucleus exhibited the phenomenon of nonlinear summation when 350-Hz and 1,700-Hz tones were presented simultaneously rather than individually; and the central, but not posterior, nucleus showed selectivity for the repetition rate of pulsed sound signals. The posterior and central thalamic nuclei also possessed distinct innervation patterns as revealed by the HRP transport patterns arising from these structures. The central nucleus was reciprocally connected with the major auditory relay stations along the frog's central auditory pathway including the superior olive, nucleus of the lateral lemniscus, and the torus semicircularis. Major projections to the lateral thalamic nucleus, ventral hypothalamus, and the telencephalic striatal complex were also observed. The posterior nucleus, on the other hand, established reciprocal connections primarily with the medial reticular nucleus, ventral midbrain tegmentum, and structures constituting of the ventral thalamic nuclei, particularly the nucleus of Bellonci. Thus, time and frequency cues contained within the species mating call, and conveying information concerning species identity, appear to be processed independently within the frog's thalamus with separate neural channels for each.

Animals↗
PubMed1987

Retina of the tadpole and frog: delayed dendritic development in a subpopulation of ganglion cells coincident with metamorphosis.

In this study, the morphology of tadpole retinal ganglion cells was compared to that of frogs to determine if changes in dendritic structure occur during metamorphosis. Ganglion cells were analyzed in the tadpole and frog after backfilling with horseradish peroxidase. Representative ganglion cells are present in the tadpole retina, which directly correspond to each of the 7 cell classes found in the frog. However, cells in 3 of these classes (1, 3, and 7) exist in morphologically immature states in retinas from tadpole stages St. XIV-XIX. New dendritic branches appear and the dendritic arbors of these ganglion cells expand during metamorphosis. We propose that the increased dendritic arborization may be followed by new synaptic contacts onto these cells, which contributes to the emergence of new physiological receptive field properties in the frog.

Animals↗
PubMed1989

Prevention of optic nerve regeneration in the frog Hyla moorei transiently delays the death of some ganglion cells.

We have previously reported that a proportion of ganglion cells die during optic nerve regeneration in the adult frog Hyla moorei (Humphrey and Beazley, '85). Here we assess the effect of preventing optic nerve regeneration on this cell loss. The optic nerve was crushed unilaterally and regeneration was allowed to progress unimpeded in one experimental series but was prevented by ligating or capping the nerve in another. We estimated total cell numbers in the ganglion cell layer from cresyl-stained wholemounts, comparing each experimental retina with its unoperated partner. At 70-78 days postcrush, mean cell numbers had fallen by 31.5% for frogs with unimpeded regeneration (N = 9), a significantly greater reduction than the 21.5% (N = 8) loss for the impeded regeneration series (p less than 0.001). Thereafter, cell numbers were stable for frogs with unimpeded regeneration. Cell death continued in the series with impeded regeneration, and losses exceeded those of frogs with unimpeded regeneration from 110 days postcrush. When regeneration was impeded, ganglion cell somas underwent an intense cell soma reaction and became arranged in rows radiating from the optic nerve head. Our findings indicate that some ganglion cells are transiently spared when regeneration of their axons is prevented. The abnormally extensive contacts formed between somas may delay ganglion cell loss. However, the eventual death of most ganglion cells shows them to be target-independent in the long term.

Animals↗
PubMed1990

Further study of the outward displacement of retinal ganglion cells during optic nerve regeneration, with a note on the normal cells of Dogiel in the adult frog.

In a previous study we observed massive retinal ganglion cell death in adult Rana pipiens after periods of optic nerve regeneration, and reported that large numbers of the surviving cells had become displaced bodily into the inner plexiform layer of the affected eye (Scalia et al.: Brain Research 344:267-280, 1985). The outwardly displaced cells could be identified as retinal ganglion cells because they could be back-filled with horseradish peroxidase (HRP) injected into the regenerated optic nerve. Quantitative observations on the abnormal outward displacement of ganglion cells are reported here. Parallel observations on normally displaced ganglion cells (cells of Dogiel) are also reported to clarify the distinctions between these two classes of cells. For the present work, injections of HRP of varying size were placed in the optic tectum bilaterally in 3 normal frogs and 9 frogs sustaining unilateral optic nerve regeneration. Most injections were centered at loci mapping the middle region of the nasal retina. The retinas were examined as flat-mounts and in-section. In 8 other frogs sustaining optic nerve regeneration, the HRP was administered bilaterally directly to the optic nerves in the orbit. Ganglion cells were labeled by retrograde transport of the HRP in the retinal ganglion cell layer in both the normal and affected eyes in areas topographically isomorphic with the tectal areas subtended by the injections. In the normal eyes, the orthotopic ganglion cells formed a strict monolayer, and virtually no cells existed in the inner plexiform layer. In the retinas sustaining optic nerve regeneration, the retinal ganglion cells abnormally displaced into the inner plexiform layer were also labeled topographically in correspondence with the injection sites. The abnormally displaced cells comprised 5.5% of the total population of surviving neurons in the retinal ganglion cell and inner plexiform layers. The mean outward dislocation of the displaced cells, as measured in one frog surviving optic nerve crush for 8 weeks, was 69.9 +/- 2.4% of the distance across the inner plexiform layer, which itself was uniformly 14.3 +/- 0.39 microns thick. Cells of Dogiel, which were embedded within the inner nuclear layer, were also labeled when the injections of HRP spread to include the area of representation of the optic disc. The labeled cells were restricted to a dorsal, peripapillary locus capping the optic disc. Therefore, some cells of Dogiel project to the tectum normally, but only from the central retina.(ABSTRACT TRUNCATED AT 400 WORDS)

Adaptation, Ocular↗
PubMed1991

Blood-nerve barrier in the frog during wallerian degeneration: are axons necessary for maintenance of barrier function?

Blood-nerve barrier tissues (endoneurial blood vessels and perineurium) of the frog's sciatic nerve were studied during chronic Wallerian degeneration to determine whether barrier function depends on the presence of intact axons. Sciatic nerves of adult frogs were transected in the abdominal cavity; the ends were tied to prevent regeneration and the distal nerve stumps were examined. Vascular permeabilities to horseradish peroxidase and to [14C]sucrose increased to day 14, returned toward normal levels by 6 weeks, and continued at near normal levels to 9 months. Perineurial permeabilities to the tracers increased by day 10 and remained elevated at 9 months. Proliferation of perineurial, endothelial, and mast cells occurred between 3 days and 6 weeks, resulting in an increased vascular space (measured with [3H]dextran) and number of vascular profiles. The perineurium increased in thickness and the mast cells increased in number. This study indicates that during Wallerian degeneration of the frog's sciatic nerve there is 1) a transitory increase in vascular permeability distal to the lesion, that is related to changes within the endoneurium; 2) an irreversible increase in permeability of the perineurium, which begins later than that seen in the endoneurial blood vessels; and 3) proliferation of non-neuronal components in the absence of regenerating neuronal elements. The results indicate that maintenance of vascular integrity does not require the presence of axons in the frog's peripheral nerve, whereas perineurial integrity and barrier function are affected irreversibly by Wallerian degeneration.

Animals↗
PubMed1995

Frog prohormone convertase PC2 mRNA has a mammalian-like expression pattern in the central nervous system and is colocalized with a subset of thyrotropin-releasing hormone-expressing neurons.

The prohormone convertase (PC2) is expressed in the mammalian central nervous system (CNS) and has been shown to play an important role in the processing of certain neuropeptide precursors and prohormones at paired basic residues. Amphibian PC2 cDNA was recently cloned for the frog Xenopus laevis, and both its sequence and its pituitary expression pattern were shown to be very similar to those of mammalian PC2. To investigate further the function of PC2 in the vertebrate CNS, we used in situ hybridization histochemistry to localize the distribution of cells expressing PC2 mRNA in the frog brain and the spinal cord. The distribution of PC2-expressing cells was also compared with that of cells expressing thyrotropin-releasing hormone (TRH) mRNA or peptide. PC2-expressing cells were detected in specific nuclei that were widely distributed in the frog CNS. In forebrain, telencephalic PC2 mRNA was found in the olfactory bulb, pallium, striatum, amygdala, and septum, and diencephalic PC2 mRNA was seen in the preoptic area, thalamus, and hypothalamus. More posteriorly, PC2 cells were localized to midbrain tegmentum, the torus semicircularis, and the optic tectum, as well as the cerebellum, brainstem, and spinal cord. Despite this wide distribution steady-state levels of PC2 mRNA were clearly different in various brain nuclei. Regions with higher levels showed good correspondence to areas shown by others in frog to contain large numbers of neuropeptide-expressing cells, including TRH cells. On the other hand, not all brain areas with high levels of TRH mRNA had high levels of PC2 mRNA. Localization studies combining in situ hybridization and immunocytochemistry showed that, at least in optic tectum and brainstem, PC2 mRNA and pro-TRH peptide coexist. These findings suggest that pro-TRH is processed by PC2 in some, but possibly not all, brain regions. Thus, different converting enzymes may be involved in pro-TRH processing in different brain regions.

Animals↗
PubMed2002

Functional expression of Kir 6.1/SUR1-K(ATP) channels in frog retinal Müller glial cells.

The retinae and brains of larval and adult amphibians survive long-lasting anoxia; this finding suggests the presence of functional K(ATP) channels. We have previously shown with immunocytochemistry studies that retinal glial (Müller) cells in adult frogs express the K(ATP) channel and receptor proteins, Kir6.1 and SUR1, while retinal neurons display Kir6.2 and SUR2A/B (Skatchkov et al., 2001a: NeuroReport 12:1437-1441; Eaton et al., in press: NeuroReport). Using both immunocytochemistry and electrophysiology, we demonstrate the expression of Kir6.1/SUR1 (K(ATP)) channels in adult frog and tadpole Müller cells. Using conditions favoring the activation of K(ATP) channels (i.e., ATP- and spermine-free cytoplasm-dialyzing solution containing gluconate) in Müller cells isolated from both adult frogs and tadpoles, we demonstrate the following. First, using the patch-clamp technique in whole-cell recordings, tolbutamide, a blocker of K(ATP) channels, blocks nearly 100% of the transient and about 30% of the steady-state inward currents and depolarizes the cell membrane by 5-12 mV. Second, inside-out membrane patches display a single-channel inward current induced by gluconate (40 mM) and blocked by ATP (200 microM) at the cytoplasmic side. The channels apparently show two sublevels (each of approximately 27-32 pS) with a total of 85-pS maximal conductance at -80 mV; the open probability follows a two-exponential mechanism. Thus, functional K(ATP) channels, composed of Kir6.1/SUR1, are present in frog Müller cells and contribute a significant part to the whole-cell K+ inward currents in the absence of ATP. Other inwardly rectifying channels, such as Kir4.1 or Kir2.1, may mediate the remaining currents. K(ATP) channels may help maintain glial cell functions during ATP deficiency.

ATP-Binding Cassette Transporters↗
PubMed2002

Application of frog (Rana tigerina Daudin) skin collagen as a novel substrate in cell culture.

Collagen and collagen-based materials have extensive application in biomedical devices and tissue engineering. The current paper pertains to the application of frog (Rana tigerina Daudin) skin collagen as a novel substrate in cell culture. The study deals with the behavior, morphology, and physiology of keratinocytes and fibroblasts over dry and reconstituted collagen substratum, which are the key cells involved in wound repair. The advantage of using frog skin collagen as a substratum lies in the ease with which the reconstituted gel can be formed. Further, frog skin collagen is highly hydrophilic, which may be attributed to the fact that amphibians, as the first vertebrates connecting water and land, must have evolved certain physiologic specializations. These studies also contribute to the hypothesis that part of the healing efficacy of frog skin may be due to the collagen since proliferation, migration, and differentiation of epithelial cells are prime requisites for a normal healing mechanism.

Animals↗
PubMed2005

D-aspartate modulates transcriptional activity in Harderian gland of frog, Rana esculenta: Morphological and molecular evidence.

In the green frog, Rana esculenta, a substantial amount of D-aspartate (D-Asp) is found endogenously within the Harderian gland (HG) following its synthesis from L-aspartate (L-Asp) by an aspartate racemase. The frog HG is an orbital seromucoid gland that displays seasonal changes in secretory activity. Our in vivo experiments, consisting of i.p. injection of 2.0 mumol/g b.w. D-Asp in frogs collected during two periods of differing glandular activity (high or medium-low secretory activity), revealed that HG can to take up and accumulate D-Asp and that this amino acid may modulate the exocrine secretion through a kinase pathway. At a time when the gland shows relatively low secretory activity, i.p. administration of D-Asp rapidly induced activation of ERK1 and an increase in cells active in RNA synthesis. This increase in transcriptional activity was followed by a significant increase in mucous secretion. By contrast, administration of exogenous D-Asp when HG was showing high activity rapidly induced inhibition of both ERK1 and transcriptional activity. Since D-Asp is known to be recognized by receptors for N-methyl-D-aspartic acid (NMDA), it is possible that in the HG, D-Asp mediated NMDA activation may enhance the kinase pathway. The above activation of opposing stimulatory and inhibitory processes could reflect different levels of NMDA-receptor activity, which could vary as a function of the level of gland activity. This study provides the first evidence of a role for this excitatory amino acid in exocrine secretion. The effects of D-Asp in HG appear to be specific since they were not seen in frogs treated with other D- or L-amino acids with known excitatory effects on neurosecretion.

Animals↗
PubMed1991

The endothelial vesicle system in cryofixed frog mesenteric capillaries analysed by ultrathin serial sectioning.

Conventional EM sections of chemically fixed capillary endothelial cells reveal numerous apparently free smooth plasmalemmal vesicles. However, the method of ultrathin (less than 150 A) serial sectioning has shown that the smooth vesicle profiles arise merely as a result of the EM thin sectioning of two sets of complex vesicular invaginations from the luminal and abluminal cell surfaces, which end blindly in the cytoplasm. While 50-70% of the total population of vesicular profiles appear to lack connections to the cell surface in conventional (500-700 A thick) EM thin sections less than 1% truly free vesicles can be found by the ultrathin serial section analyses. In the present study it is examined whether similar conclusions apply to endothelial cells which were directly frozen by slam-freezing and subsequently freeze-substituted. The three-dimensional organization of the plasmalemmal vesicular system was analyzed in four series of 19, 18, 13, and 10 ultrathin sections (approximately 110 A thick) of capillaries from frog mesenteries quickly excised from decapitated frogs (Rana pipiens). None of 920 vesicular profiles (diameter 500-1,200 A) which appeared free in individual thin sections of the series represented free vesicles; all profiles either communicated with other vesicles, the cell surface, or in rare cases turned out to be part of cytoplasmic tubular membrane structures. It is concluded that free smooth plasmalemmal vesicles are very rare in rapidly frozen as well as in directly fixed frog capillary endothelium. The volume density of profiles (13-15%), the proportion of apparently free vesicle profiles (70%), and interconnected profiles (20%) were similar to the picture previously found in single EM sections of frog mesenteric capillaries. No transendothelial channels were found in the four series of ultrathin sections of capillaries. However, continuities between the luminal and abluminal cell surfaces were seen in the endothelium of venules. Furthermore, in the ultrathin series of the capillaries, vesicular units belonging to the two sets of invaginations and cytoplasmic tubular membrane structures were in more cases found in very close contact-as fused to share one unit membrane. If this finding is representative for the in vivo situation, it may reflect that the vesicular system represents a highly dynamic system with possibilities for mixing of membranes, cellular traffic of lipid, membrane proteins, and receptors between internal compartments and the cell surfaces, as well as occasional exchange of macromolecules between blood and tissue through rare temporary connections between the two sets of surface invaginations, without actually moving vesicles.

Animals↗
PubMed2001

Immunohistochemical distribution of enkephalin, substance P, and somatostatin in the brainstem of the leopard frog, Rana pipiens.

The brainstems of frogs contain many of the neurochemicals that are found in mammals. However, the clustering of nuclei near the ventricles makes it difficult to distinguish individual cell groups. We addressed this problem by combining immunohistochemistry with tract tracing and an analysis of cell morphology to localize neuropeptides within the brainstem of Rana pipiens. We injected a retrograde tracer, Fluoro-Gold, into the spinal cord, and, in the same frog, processed adjacent sections for immunohistochemical location of antibodies to the neuropeptides enkephalin (ENK), substance P (SP), and somatostatin (SOM). SOM+ cells were more widespread than cells containing immunoreactivity (ir) to the other substances. Most reticular nuclei in frog brainstem contained ir to at least one of these chemicals. Cells with SOM ir were found in nucleus (n.) reticularis pontis oralis, n. reticularis magnocellularis, n. reticularis paragigantocellularis, n. reticularis dorsalis, the optic tectum, n. interpeduncularis, and n. solitarius. ENK-containing cell bodies were found in n. reticularis pontis oralis, n. reticularis dorsalis, the nucleus of the solitary tract, and the tectum. The midbrain contained most of the SP+ cells. Six nonreticular nuclei (griseum centrale rhombencephali, n. isthmi, n. profundus mesencephali, n. interpeduncularis, torus semicircularis laminaris, and the tectum) contained ir to one or more of the substances but did not project to the spinal cord. The descending tract of V, and the rubrospinal, reticulospinal, and solitary tracts contained all three peptides as did the n. profundus mesencephali, n. isthmi, and specific tectal layers. Because the distribution of neurochemicals within the frog brainstem is similar to that of amniotes, our results emphasize the large amount of conservation of structure, biochemistry, and possibly function that has occurred in the brainstem, and especially in the phylogenetically old reticular formation.

Animals↗
PubMed2001

Expression of FTZ-F1alpha in frog testicular cells.

Fushi tarazu transcription factor-1 (FTZ-F1), a member of a nuclear hormone receptor superfamily, is a transcriptional regulator for fushi tarazu gene expression in Drosophila (Ueda et al., '90). We have cloned a homologue (rrFTZ-F1alpha) of the FTZ-F1 gene of the frog Rana rugosa. The gene, in frogs, has been shown to have high expression level in the testis (Nakajima et al., 2000). In this study, the RT-PCR analysis showed that the FTZ-F1alpha mRNA level in adult frogs did not change throughout the year, even during hibernation. However, when immunohistological studies using the anti-rrFTZ-F1alpha antibody were employed to examine which testicular cells expressed this gene, Sertoli cells were found to produce rrFTZ-F1alpha in the two seasons: the breeding season (from March through May) and the pre-hibernating season (from October through November). Interstitial cells, however, did it in only the breeding season (from April through May). Taken together, the results suggest that the rrFTZ-F1alpha expression is regulated at the post-transcriptional step, and that the rrFTZ-F1alpha may play an important role(s) in the seasonal activities of Sertoli and interstitial cells in the frog testis. J. Exp. Zool. 290:182-189, 2001.

Animals↗
PubMed1986

The cellular basis of the calcium dependence of GnRH-stimulated gonadotropin release from frog, Rana pipiens, pituitaries.

An in vitro superfusion system was used in an attempt to identify the cellular systems involved in the Ca2+ dependence of gonadotropin-releasing hormone (GnRH) actions in the frog, Rana pipiens. Superfusion with 5 microM A23187 (a calcium ionophore) or phorbol myristate acetate (an analog of diacylglycerides) caused marked increases in luteinizing hormone (LH) and follicle-stimulating hormone (FSH) secretion. Exclusion of Ca2+ from the medium prevented the stimulatory effects of PMA. The potent stimulator of adenylate cyclase, forskolin, caused only slight stimulation of LH and FSH secretion, which was also prevented by removal of Ca2+. The cytoskeletal disruptive agents colchicine, nocodazole, and cytochalasin B, and the calmodulin inhibitors, trifluoperazine and pimozide, had no significant effects on the action of GnRH. Overall, these results indicate that the major mechanisms of Ca2+ involvement in the response to GnRH by the frog pituitary are similar to that of mammals, with the possible exceptions of lesser roles for calmodulin and the cytoskeleton in the frog. The study suggests that polyphosphatidyl-inositol-diacylglyceride metabolism may be critical in understanding the mechanism of GnRH action in frogs.

Animals↗
PubMed1987

Mechanisms regulating the ovarian cycle in the frog Rana cyanophlyctis: effect of unilateral ovariectomy, season, feeding, human chorionic gonadotrophin, and estradiol-17 beta.

Effects of 30-day unilateral ovariectomy (ULO) on compensatory ovarian hypertrophy (COH) was studied in adult frog Rana cyanophlyctis in relation to reproductive phase/season, feeding, and treatment with human chorionic gonadotrophin (HCG) or estradiol-17 beta. Compensatory growth of the remaining ovary was assessed in terms of weight as well as changes in the dynamics of the pool sizes of different oocytes viz., first growth phase (FGP), medium-sized second growth phase (MSGP), large-sized second growth phase (LSGP), and atretic oocytes. The frogs were fed with live guppies 6 days a week. The effect of underfeeding on COH was studied in the frogs fed once a week. The ovaries removed at operation and those of the sham-operated frogs were used for comparison (controls). COH occurred in both pre- and postbreeding phases (February and November, respectively). In both instances follicular atresia was greatly reduced. In February, COH was due to recruitment of both MSGP and LSGP oocytes. The number of these oocytes increased significantly over controls, and the ovarian weight nearly doubled. However, in November COH occurred because of an increase in FGP oocytes and therefore total oocytes, but there were no changes in the ovarian weights. Administration of 20 IU HCG (6 days a week) had no influence on the basic pattern of COH-response exhibited by the remaining ovary in relation to reproductive phase/season, but it increased the recruitment of oocytes, i.e., FGP oocytes in November and SGP oocytes in February. Follicular atresia was drastically reduced with HCG. Underfeeding or treatment with estradiol-17 beta abolished the COH in both February and November.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1991

Protective effects of manganese, cobalt, nickel, and barium against a calcium paradox in the isolated frog heart.

The effect of inorganic slow channel blockers on the calcium paradox in the frog heart was examined. Addition of the divalent cations of manganese, cobalt, nickel, or barium during calcium depletion protected the frog heart against a calcium paradox. This protective effect was indicated by reduced protein release, maintenance of electrical activity, and recovery of mechanical activity during reperfusion. Tissue calcium determination results showed that in the control paradox in the absence of divalent cations, there is an efflux of calcium from myocardial cells during calcium depletion and a massive influx of calcium during the following reperfusion, leading to a calcium overload. Divalent cations protected frog myocardial cells, when present in the calcium-free perfusion medium, by reducing both calcium efflux during calcium depletion and the massive calcium influx during reperfusion. The effectiveness of the added divalent cations showed a strong dependence upon their ionic radius. The most potent inhibitors of the calcium paradox in the frog heart were the divalent cations having an ionic radius closer to the ionic radius of calcium. These results are discussed in terms of the possible mechanism involved in the protective effect of manganese, cobalt, nickel, and barium.

Animals↗
PubMed1993

On the existence of epithelial chloride channels in intact frogs and crayfish.

Chloride channels in isolated amphibian skins have been shown to be open (high PCl) when the transepithelial potential (TEP) across the skin is > 30 mV, inside positive, and [Cl-] in the external medium is high. In the absence of either of these signals PCl is much lower. Using Cl- efflux as a criterion, it is shown here that when the same cues are presented to intact frogs, channels open as in the in vitro preparation. The augmented efflux is partially inhibited by high I- in both preparations. However, salt-depleted frogs do not show channel opening, differing in this regard from isolated skins. Elevating external [Cl-] resulted in an even larger efflux stimulation in crayfish than in frogs. The TEP in these animals is usually small and sometimes blood negative; it does not appear to play a role in augmenting the efflux. Gluconate and NO3- also caused Cl efflux to increase markedly. Salt-depletion reduced the stimulation by high [Cl-] but did not abolish it as in frogs. Iodide had no effect on chloride efflux in crayfish. The results may indicate that gated channels exist in these animals, but interpretation is complicated by the possibility that the augmented efflux is due to exchange diffusion.

Animals↗