Cryoprotective effects of longan honey on preantral follicle integrity of rat ovary post vitrification.

BACKGROUND: Longan honey (LH) has the potential as a natural extracellular cryoprotectant to maintain the integrity of intact preantral follicles against the cryoinjury during ovarian vitrification. OBJECTIVE: This research determined the cryoprotective effects of logan honey on preantral follicles integrity of rat ovary post-vitrification. MATERIALS AND METHODS: After vitrification, the follicle index was determined by observing the ovarian histological sections made using the paraffin method with hematoxylin-eosin staining and analyzed using Optilab 3.0 and Image Raster software. RESULTS: The results showed that the combination of ethylene glycol (EG) with LH and a dose variation of 7.5 %-15 % (KP1-KP4) increased the density of follicles, the number of intact follicles in G2 and G3, with a decrease in the atretic follicles in G1 better compared to the use of EG only (KKP1-KKP2). The differences in the histological structur e of preantral follicles affected the doses of LH needed by each type of follicle to maintain the integrity of the follicular structure from cryoinjury effects. The comparison of the G2 total follicle index values were KKP1 (90.7 ± 18.3), KKP2 (115.6 ± 9.9) < KP1 (193.6 ± 10.7), KP2 (189.3 ± 10.5), KP3 (182.2 ± 27.1) and KP4 (169.4±8.8). Among the variations in the dose of LH 7.5 % - 15%, the highest increase in the G3 index value was found in primary (51.7 ± 9.8), tertiary (43.1 ± 8.8), secondary (33.9 ± 4.7), and primordial (28.7 ± 2.5) follicles of KP3 (7.5 % of LH). CONCLUSION: The primary and tertiary follicular stages maintain the best integrity and can be used after the vitrification of rat ovaries. doi.org/10.54680/fr23110110712.

Expression of Ca2+/calmodulin-dependent protein kinase (CaMK) Ibeta2 in developing rat CNS.

We observed the onset time and distribution pattern of beta2 isoform of Ca2+/calmodulin-dependent protein kinase I (CaMKIbeta2) in the CNS of the rat during the embryonic period until birth using an immunohistochemical method. The expression of CaMKIbeta2 started at embryological day 10 when the three primary brain vesicles and neural tube are generated from the neural plate. During the embryonic period, highly immunoreactive products were ubiquitously detected in neurons in the CNS, although neurons in the caudate-putamen and globus pallidus were faintly immunostained or immunonegative. High expression of CaMKIbeta2 persisted in the olfactory bulb, lymbic system, neocortex, septal nuclei, amygdala complex, some hypothalamic nuclei, pontine nuclei, Purkinje cells and granule cells in the cerebellar cortex through the developing period. At the subcellular level, CaMKIbeta2 was strongly expressed in nuclei of neurons but faintly in their cytoplasm, suggesting that this protein has an important role in the nuclear signaling pathway. This study demonstrates that expression of CaMKIbeta2 begins at the earliest developmental stage of the rat CNS and persists through the developing period.

Distribution of Ca(2+)/calmodulin-dependent protein kinase I beta 2 in the central nervous system of the rat.

Recently, we reported the mRNA localization of Ca(2+)/calmodulin-dependent protein kinase I beta 2 isoform (CaMKIbeta2) in the mouse nervous system. In the present study, polyclonal antibody against CaMKIbeta2 was generated and used to investigate the distribution of the enzyme within the central nervous system of the rat. Interestingly some differences were observed between the enzyme localization and previous mRNA detection [J. Neurochem. 268 (1999) 26512]. The strongest expression of the enzyme was found in pontine nuclei. Immunopositive fibers could be traced through the middle cerebellar peduncle until they reached the cerebellum. Quite strong staining could also be observed in almost all of the neurons in the neocortex, hippocampus, amygdala, hypothalamus, brainstem and cerebellum, including the nuclei of the cranial nerves and Purkinje cell layer of the cerebellar cortex which was not clearly detected in the previous in situ hybridization study. In the spinal cord, CaMKIbeta2 could be detected in the gray matter with stronger expression in the dorsal horn. CaMKIbeta2 showed very strong nuclear localization but was also present in the cytoplasm of some neurons. Such localization suggests that CaMKIbeta2 may be involved in many neuronal functions in the central nervous system, including the possibility of important roles in nuclear signal transduction.

Vesicourethral sphincter dysfunction in ncx deficient mice with an increased neuronal cell number in vesical ganglia.

PURPOSE: Ncx/Hox11L.1 knockout mice have a megacolon with an increased number of neuronal cells in the enteric ganglia. Since Ncx/Hox11L.1 is expressed in neuronal cells in the vesical ganglia, we examined lower urinary tract function and the number of neuronal cells in the vesical ganglia in Ncx/Hox11L.1 knockout mice. METHODS: Female knockout and control mice were investigated in regard to voiding frequency, and cystometry and histological studies were done. The number of neuronal cells in the vesical ganglia was observed by staining with nicotinamide adenine dinucleotide phosphate diaphorase and cuprolinic blue. RESULTS: In knockout mice voiding frequency was 2-fold and bladder capacity was less than in controls. Although bladder structure was histologically similar in knockout mice and controls, cystometry showed that threshold and remaining pressure was less in knockout mice. Neuronal cells positive for nicotinamide adenine dinucleotide phosphate diaphorase or cuprolinic blue were more numerous in the vesical ganglia of knockout mice than controls. The intraperitoneal injection of a nitric oxide synthase inhibitor increased threshold and remaining pressure on cystometry in knockout mice to the control level. CONCLUSIONS: The increased number of neuronal cells in the vesical ganglia induces vesicourethral sphincter muscle dysfunction in knockout mice. Since administering a nitric oxide synthase inhibitor somewhat overcomes the dysfunction, the amount of nitric oxide in vesical nerve cells is important for controlling vesicourethral sphincter muscle function.