Immunodetection of Tau microtubule-associated protein in human sperm and testis

Sigala, Julien; Jumeau, Fanny; Caillet-Boudin, Marie-Laure; Sergeant, Nicolas; Ballot, Caroline; Rigot, Jean-Marc; Marcelli, F.; Tardivel, Meryem; Buée, Luc; Mitchell, Valérie

doi:10.4103/1008-682x.136446

Cited by 8 publications

(3 citation statements)

References 9 publications

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“…The 2 kb transcript was found to code for a major nuclear species of Tau [ 71 ] and has also been reported in the human frontal cortex by Michel Goedert [ 19 ] and in testicular spermatid manchette [ 72 ]. The presence of Tau in the sperm and testis has also been reported independently [ 73 , 74 ]. It is not clear whether the isoform-specific distribution of Tau to either the nucleus, soma and axons reported in the murine brain [ 44 ] is dictated by different transcripts (2 kb and 6 kb), or whether analogous transcripts exist in other species e.g.…”

Section: Atypical/non-standard Functions Of Taumentioning

confidence: 63%

Atypical, non-standard functions of the microtubule associated Tau protein

Sotiropoulos

Galas

Silva

et al. 2017

acta neuropathol commun

159

134

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Since the discovery of the microtubule-associated protein Tau (MAPT) over 40 years ago, most studies have focused on Tau’s role in microtubule stability and regulation, as well as on the neuropathological consequences of Tau hyperphosphorylation and aggregation in Alzheimer’s disease (AD) brains. In recent years, however, research efforts identified new interaction partners and different sub-cellular localizations for Tau suggesting additional roles beyond its standard function as microtubule regulating protein. Moreover, despite the increasing research focus on AD over the last decades, Tau was only recently considered as a promising therapeutic target for the treatment and prevention of AD as well as for neurological pathologies beyond AD e.g. epilepsy, excitotoxicity, and environmental stress. This review will focus on atypical, non-standard roles of Tau on neuronal function and dysfunction in AD and other neurological pathologies providing novel insights about neuroplastic and neuropathological implications of Tau in both the central and the peripheral nervous system.

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Section: Atypical/non-standard Functions Of Taumentioning

confidence: 63%

Atypical, non-standard functions of the microtubule associated Tau protein

Sotiropoulos

Galas

Silva

et al. 2017

acta neuropathol commun

159

134

View full text Add to dashboard Cite

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“…Originally thought to be a neuronal-specific protein, Tau expression has now been demonstrated in oligodendrocytes, astrocytes and various retinal cell types in the central nervous system (CNS), and has also been reported in many other tissues, including the peripheral nervous system (PNS), submandibular gland, tonsil, liver, colon, heart, skeletal muscle, breast, kidney, skin, pancreas, adrenal gland, reproductive organs and salivary glands [18][19][20][21][22][23][24][25][26][27][28][29][30][31][32][33][34]. Given its pathological relevance, studies to date have focussed largely on the role of Tau hyperphosphorylation and aggregation in driving neurodegeneration, mostly employing overexpression models or patient brain samples where Tau is expressed at high levels and is often found in aggregated states.…”

Section: Introductionmentioning

confidence: 99%

Validation of Tau Antibodies for Use in Western Blotting and Immunohistochemistry

Ellis

Lekka

Tulmin

et al. 2023

Preprint

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BackgroundThe microtubule-associated protein Tau has attracted diverse and increasing research interest, with Tau being mentioned in the title/abstract of nearly 34,000 PubMed-indexed publications to date. To accelerate studies into Tau biology, the characterisation of its multiple proteoforms, including disease-relevant post-translational modifications (PTMs), and its role in neurodegeneration, a multitude of Tau-targeting antibodies have been developed, with hundreds of distinct antibody clones currently available for purchase. Nonetheless, concerns over antibody specificity and limited understanding of the performance of many of these reagents has hindered research.MethodsWe have employed a range of techniques in combination with samples of murine and human origin to characterise the performance and specificity of 53 commercially-available Tau antibodies by Western blot, and a subset of these, 35 antibodies, in immunohistochemistry.ResultsContinued expression of residual protein was found in presumptive Tau “knockout” human cells and further confirmed through mass-spectrometry proteomics, providing evidence of Tau isoforms generated by exon skipping. Importantly, many total and isoform-specific antibodies failed to detect this residual Tau, as well as Tau expressed at low, endogenous levels, thus highlighting the importance of antibody choice. Our data further reveal that the binding of several “total” Tau antibodies, which are assumed to detect Tau independently of post-translational modifications, was partially inhibited by phosphorylation. Many antibodies also displayed non-specific cross-reactivity, with some total and phospho-Tau antibodies cross-reacting with MAP2 isoforms, while the “oligomer-specific” T22 antibody detected monomeric Tau on Western blot. Regardless of their specificity, with one exception, the phospho-Tau antibodies tested were found to not detect the unphosphorylated protein.ConclusionsWe identify Tau antibodies across all categories (total, PTM-dependent and isoform-specific) that can be employed in Western blot and/or immunohistochemistry applications to reliably detect even low levels of Tau expression with high specificity. This is of particular importance for studying Tau in non-neuronal cells and peripheral tissues, as well as for the confident validation of knockout cells and/or animal models. This work represents an extensive resource that serves as a point of reference for future studies. Our findings may also aid in the re-interpretation of existing data and improve reproducibility of Tau research.

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“…Humanized TTBK1 transgenic mice display phosphorylated neurofilament aggregation and age-dependent memory impairment (Sato et al , 2008), whereas TTBK2 mutant mice manifest defects in primary cilia formation, loss of Sonic hedgehog signaling activity, and ultimately die during embryogenesis (Bouskila et al , 2011; Goetz et al , 2012). MAP2 and Tau are also expressed in the testes (Loveland et al , 1996; Inoue et al , 2014; Sigala et al , 2014). However, Tau and MAP2 are detected before development of sperm flagella and with nuclear localization during meiosis (Loveland et al , 1996, 1999; Inoue et al , 2014).…”

Section: Introductionmentioning

confidence: 99%

Tau tubulin kinase is required for spermatogenesis and development of motile cilia in planarian flatworms

Magley

Rouhana

2019

MBoC

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Cilia are microtubule-based structures that protrude from the apical surface of cells to mediate motility, transport, intracellular signaling, and environmental sensing. Tau tubulin kinases (TTBKs) destabilize microtubules by phosphorylating microtubule-associated proteins (MAPs) of the MAP2/Tau family, but also contribute to the assembly of primary cilia during embryogenesis. Expression of TTBKs is enriched in testicular tissue, but their relevance to reproductive processes is unknown. We identified six TTBK homologues in the genome of the planarian Schmidtea mediterranea ( Smed-TTBK-a, -b, -c, -d, -e, and -f), all of which are preferentially expressed in testes. Inhibition of TTBK paralogues by RNA interference (RNAi) revealed a specific requirement for Smed-TTBK-d in postmeiotic regulation of spermatogenesis. Disrupting expression of Smed-TTBK-d results in loss of spermatozoa, but not spermatids. In the soma, Smed-TTBK-d RNAi impaired the function of multiciliated epidermal cells in propelling planarian movement, as well as the osmoregulatory function of protonephridia. Decreased density and structural defects of motile cilia were observed in the epidermis of Smed-TTBK-d(RNAi) by phase contrast, immunofluorescence, and transmission electron microscopy. Altogether, these results demonstrate that members of the TTBK family of proteins are postmeiotic regulators of sperm development and also contribute to the formation of motile cilia in the soma.

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Immunodetection of Tau microtubule-associated protein in human sperm and testis

Cited by 8 publications

References 9 publications

Atypical, non-standard functions of the microtubule associated Tau protein

Atypical, non-standard functions of the microtubule associated Tau protein

Validation of Tau Antibodies for Use in Western Blotting and Immunohistochemistry

Tau tubulin kinase is required for spermatogenesis and development of motile cilia in planarian flatworms

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