https://nova.newcastle.edu.au/vital/access/manager/Index ${session.getAttribute("locale")} 5 https://nova.newcastle.edu.au/vital/access/manager/Repository/uon:44975 In vivo, the inner ear is difficult to study due to the osseous nature of the otic capsule and its encapsulation within an intricate bony labyrinth. As such, mammalian inner ear explants are an invaluable tool for the study and manipulation of the complex intercellular connections, structures, and cell types within this specialised organ. The greatest strength of this technique is that the complete organ of Corti, or peripheral vestibular organs including hair cells, supporting cells and accompanying neurons, is maintained in its in situ form. The greatest weakness of in vitro hair cell preparations is the short time frame in which the explanted tissue remains viable. Yet, cochlear explants have proven to be an excellent experimental model for understanding the fundamental aspects of auditory biology, substantiated by their use for over 40 years. In this protocol, we present a modernised inner ear explant technique that employs organotypic cell culture inserts and serum free media. This approach decreases the likelihood of explant damage by eliminating the need for adhesive substances. Serum free media also restricts excessive cellular outgrowth and inter-experimental variability, both of which are side effects of exogenous serum addition to cell cultures. The protocol described can be applied to culture both cochlear and vestibular explants from various mammals. Example outcomes are demonstrated by immunohistochemistry, hair cell quantification, and electrophysiological recordings to validate the versatility and viability of the protocol.]]> Wed 26 Oct 2022 08:46:25 AEDT ]]> https://nova.newcastle.edu.au/vital/access/manager/Repository/uon:6547 Wed 11 Apr 2018 15:10:36 AEST ]]> https://nova.newcastle.edu.au/vital/access/manager/Repository/uon:1306 50%) compared with controls for each mutant. mIPSC decay times were unchanged in spa/spa (4.5 ± 0.3 vs 4.7 ± 0.2 ms), reduced in spd/spd (2.7 ± 0.2 vs 4.7 ± 0.2 ms), and increased in ot/ot (12.3 ± 1.2 vs 4.8 ± 0.2 ms). Thus, in spastic, GlyRs are functionally normal but reduced in number, whereas in spasmodic, GlyR kinetics is faster. The oscillator mutation results in complete absence of α1-containing GlyRs; however, some non-α1-containing GlyRs persist at synapses. Fluctuation analysis of membrane current, induced by glycine application to outside-out patches, showed that mean single-channel conductance was increased in spa/spa (64.2 ± 4.9 vs 36.1 ± 1.4 pS), but unchanged in spd/spd (32.4 ± 2.1 vs 35.3 ± 2.1 pS). GlyR-mediated whole-cell currents in spa/spa exhibited increased picrotoxin sensitivity (27 vs 71% block for 100 µM), indicating α1 homomeric GlyR expression. The picrotoxin sensitivity of evoked glycinergic IPSCs and conductance of synaptic GlyRs, as determined by nonstationary variance analysis, were identical for spa/spa and controls. Together, these findings show the three mutations disrupt GlyR-mediated inhibition via different physiological mechanisms, and the spastic mutation results in "compensatory" α1 homomeric GlyRs at extrasynaptic loci.]]> Wed 11 Apr 2018 14:28:46 AEST ]]> https://nova.newcastle.edu.au/vital/access/manager/Repository/uon:21342 Wed 11 Apr 2018 12:17:52 AEST ]]> https://nova.newcastle.edu.au/vital/access/manager/Repository/uon:6543 Wed 11 Apr 2018 12:03:05 AEST ]]> https://nova.newcastle.edu.au/vital/access/manager/Repository/uon:3122 Wed 11 Apr 2018 10:52:30 AEST ]]> https://nova.newcastle.edu.au/vital/access/manager/Repository/uon:7526 Wed 11 Apr 2018 10:11:50 AEST ]]> https://nova.newcastle.edu.au/vital/access/manager/Repository/uon:46022 300 inflammatory genes were similarly regulated in the uterine endometrium by mating independently of paternal diet, and 13 were dysregulated by HFD-fed compared with CD-fed males. Seminal vesicle fluid factors reduced in HFD-fed males, including TGF-β1, IL-10, and TNF, were among the predicted upstream regulators of differentially regulated genes. Additionally, the T-cell response induced by mating with CD-fed males was blunted after mating with HFD-fed males, with 27% fewer CD4⁺ T cells, 26% fewer FOXP3⁺CD4⁺ regulatory T cells (Treg) cells, and 19% fewer CTLA4⁺ Treg cells, particularly within the NRP1⁺ thymic Treg cell population. These findings demonstrate that an obesogenic HFD alters the composition of seminal vesicle fluid and impairs seminal plasma capacity to elicit a favorable pro-tolerogenic immune response in females at conception.]]> Wed 09 Nov 2022 11:05:03 AEDT ]]> https://nova.newcastle.edu.au/vital/access/manager/Repository/uon:38997 Tue 05 Apr 2022 14:09:29 AEST ]]> https://nova.newcastle.edu.au/vital/access/manager/Repository/uon:53503 Thu 30 Nov 2023 15:57:23 AEDT ]]> https://nova.newcastle.edu.au/vital/access/manager/Repository/uon:45401 Thu 27 Oct 2022 17:29:15 AEDT ]]> https://nova.newcastle.edu.au/vital/access/manager/Repository/uon:1255 Thu 09 Nov 2023 15:42:12 AEDT ]]> https://nova.newcastle.edu.au/vital/access/manager/Repository/uon:7528 Sat 24 Mar 2018 08:38:31 AEDT ]]> https://nova.newcastle.edu.au/vital/access/manager/Repository/uon:7337 Sat 24 Mar 2018 08:35:13 AEDT ]]> https://nova.newcastle.edu.au/vital/access/manager/Repository/uon:11329 Sat 24 Mar 2018 08:12:34 AEDT ]]> https://nova.newcastle.edu.au/vital/access/manager/Repository/uon:10807 Sat 24 Mar 2018 08:11:50 AEDT ]]> https://nova.newcastle.edu.au/vital/access/manager/Repository/uon:21158 FZR1 activity in the male germline led to both a mitotic and a meiotic testicular defect resulting in infertility due to the absence of mature spermatozoa. Spermatogonia in the prepubertal testes of such mice had abnormal proliferation and delayed entry into meiosis. Although early recombination events were initiated, male germ cells failed to progress beyond zygotene and underwent apoptosis. Loss of APC/C^FZR1 activity was associated with raised cyclin B1 levels, suggesting that CDK1 may trigger apoptosis. By contrast, female FZR1Δ mice were subfertile, with premature onset of ovarian failure by 5 months of age. Germ cell loss occurred embryonically in the ovary, around the time of the zygotene-pachytene transition, similar to that observed in males. In addition, the transition of primordial follicles into the growing follicle pool in the neonatal ovary was abnormal, such that the primordial follicles were prematurely depleted. We conclude that APC/C^FZR1 is an essential regulator of spermatogonial proliferation and early meiotic prophase I in both male and female germ cells and is therefore important in establishing the reproductive health of adult male and female mammals.]]> Sat 24 Mar 2018 08:00:18 AEDT ]]> https://nova.newcastle.edu.au/vital/access/manager/Repository/uon:16896 Sat 24 Mar 2018 07:58:48 AEDT ]]> https://nova.newcastle.edu.au/vital/access/manager/Repository/uon:6573 Sat 24 Mar 2018 07:49:17 AEDT ]]> https://nova.newcastle.edu.au/vital/access/manager/Repository/uon:6570 Sat 24 Mar 2018 07:49:16 AEDT ]]> https://nova.newcastle.edu.au/vital/access/manager/Repository/uon:6577 Sat 24 Mar 2018 07:49:16 AEDT ]]> https://nova.newcastle.edu.au/vital/access/manager/Repository/uon:6576 Sat 24 Mar 2018 07:49:16 AEDT ]]> https://nova.newcastle.edu.au/vital/access/manager/Repository/uon:6586 Sat 24 Mar 2018 07:49:07 AEDT ]]> https://nova.newcastle.edu.au/vital/access/manager/Repository/uon:6563 Sat 24 Mar 2018 07:47:07 AEDT ]]> https://nova.newcastle.edu.au/vital/access/manager/Repository/uon:6564 Sat 24 Mar 2018 07:47:07 AEDT ]]> https://nova.newcastle.edu.au/vital/access/manager/Repository/uon:132 Sat 24 Mar 2018 07:43:12 AEDT ]]> https://nova.newcastle.edu.au/vital/access/manager/Repository/uon:276 30% of mutations seem to cluster on proximal Xp and in the pericentric region. In a systematic screen of brain-expressed genes from this region in 210 families with XLMR, we identified seven different mutations in JARID1C, including one frameshift mutation and two nonsense mutations that introduce premature stop codons, as well as four missense mutations that alter evolutionarily conserved amino acids. In two of these families, expression studies revealed the almost complete absence of the mutated JARID1C transcript, suggesting that the phenotype in these families results from functional loss of the JARID1C protein. JARID1C (Jumonji AT-rich interactive domain 1C), formerly known as "SMCX," is highly similar to the Y-chromosomal gene JARID1D/SMCY, which encodes the H-Y antigen. The JARID1C protein belongs to the highly conserved ARID protein family. It contains several DNA-binding motifs that link it to transcriptional regulation and chromatin remodeling, processes that are defective in various other forms of mental retardation. Our results suggest that JARID1C mutations are a relatively common cause of XLMR and that this gene might play an important role in human brain function.]]> Sat 24 Mar 2018 07:43:03 AEDT ]]> https://nova.newcastle.edu.au/vital/access/manager/Repository/uon:22273 IN), resting membrane potential, AP amplitude, half-width and AHP amplitude were similar across spinal cord regions in both neonates and adults (~100 neurons for each region and age). In contrast, these intrinsic membrane properties differed dramatically between neonates and adults. Five types of AP discharge were observed during depolarizing current injection. In neonates, single spiking dominated (~40%) and the proportions of each discharge category did not differ across spinal regions. In adults, initial bursting dominated in each spinal region, but was significantly more prevalent in rostral segments (49% of neurons in C2-4 vs. 29% in L3-5). During development the dominant AP discharge pattern changed from single spiking to initial bursting. The rapid A-type potassium current (I_Ar) dominated in neonates and adults, but its prevalence decreased (~80% vs. ~50% of neurons) in all regions during development. I_Ar steady state inactivation and activation also changed in upper cervical and lumbar regions during development. Together, our data show the intrinsic properties of SDH neurons are generally conserved in the three spinal cord regions examined in both neonate and adult mice. We propose the conserved intrinsic membrane properties of SDH neurons along the length of the spinal cord cannot explain the marked differences in pain experienced in the limbs, viscera, and head and neck.]]> Sat 24 Mar 2018 07:17:39 AEDT ]]> https://nova.newcastle.edu.au/vital/access/manager/Repository/uon:47486 Mon 23 Jan 2023 11:47:26 AEDT ]]> https://nova.newcastle.edu.au/vital/access/manager/Repository/uon:47962 Mon 13 Feb 2023 15:03:17 AEDT ]]> https://nova.newcastle.edu.au/vital/access/manager/Repository/uon:47756 Nes^-/-) mice. We found that the proliferation of Nes^-/- neural stem cells was not altered, but neurogenesis in the hippocampal dentate gyrus of Nes^-/- mice was increased. Surprisingly, the proneurogenic effect of nestin deficiency was mediated by its function in the astrocyte niche. Through its role in Notch signaling from astrocytes to neural stem cells, nestin negatively regulates neuronal differentiation and survival; however, its expression in neural stem cells is not required for normal neurogenesis. In behavioral studies, nestin deficiency in mice did not affect associative learning but was associated with impaired long-term memory.]]> Fri 27 Jan 2023 10:25:04 AEDT ]]> https://nova.newcastle.edu.au/vital/access/manager/Repository/uon:48979 Fri 21 Apr 2023 09:29:59 AEST ]]> https://nova.newcastle.edu.au/vital/access/manager/Repository/uon:32098 Fri 03 Dec 2021 10:35:09 AEDT ]]>