Prion protein is necessary for normal synaptic function

Collinge, John; Whittington, Miles A.; Sidle, Katie; Smith, Corinne J.; Palmer, Mark S.; Clarke, Anthony R.; Jefferys, John G. R.

doi:10.1038/370295a0

Cited by 723 publications

(453 citation statements)

References 21 publications

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“…Consistent with our observations in the PRNP −/− cattle, Prnp −/− mice with exclusive disruption on the Prnp ORF remained healthy 2,3 and showed only slightly abnormal phenotypes, such as altered synaptic function 29,30 and sleep-wake circadian rhythms 31,32 . However, the phenotype in the synaptic function appears to be normal in other murine genetic backgrounds 33 .We have monitored sleep-wake activity in the knockout cattle, along with age-, sex-and breed-matched wild-type controls, at frequent intervals throughout the day and night for one week, but did not observe any obvious alterations.…”

supporting

confidence: 90%

Production of cattle lacking prion protein

et al. 2006

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Prion diseases are caused by propagation of misfolded forms of the normal cellular prion protein PrP C , such as PrP BSE in bovine spongiform encephalopathy (BSE) in cattle and PrP CJD in Creutzfeldt-Jakob disease (CJD) in humans 1 . Disruption of PrP C expression in mice, a species that does not naturally contract prion diseases, results in no apparent developmental abnormalities [2][3][4][5] . However, the impact of ablating PrP C function in natural host species of prion diseases is unknown. Here we report the generation and characterization of PrP C -deficient cattle produced by a sequential gene-targeting system 6 . At over 20 months of age, the cattle are clinically, physiologically, histopathologically, immunologically and reproductively normal. Brain tissue homogenates are resistant to prion propagation in vitro as assessed by protein misfolding cyclic amplification 7 . PrP C -deficient cattle may be a useful model for prion research and could provide industrial bovine products free of prion proteins.To generate PrP C -deficient (PRNP −/− ) cattle, we transfected a male Holstein primary fetal fibroblast line 6594 with first and second knockout (KO) vectors (pBPrP(H)KOneo and pBPrP (H)KOpuro vectors) 6 to sequentially disrupt the two alleles of PRNP. PRNP −/− fetal cell lines were established at 40-60 d of gestation and three of the PRNP −/− fetal cell lines (5211, 5232 and 4296) were recloned to produce calves (Table 1 and Fig. 1a). To verify that the calves possess the PRNP −/− genotype, we collected ear biopsies and established fibroblast cell lines for genotyping. Genotyping was done by genomic PCR specific to each gene targeting event 6 (primer pairs: neoF7 × neoR7 and puroF14 × puroR14, Fig. 1b COMPETING INTERESTS STATEMENTThe authors declare competing financial interests (see the Nature Biotechnology website for details).Reprints and permissions information is available online at http://npg.nature.com/reprintsandpermissions/ NIH Public Access Fig. 1c) and confirmed the disruption of PRNP-specific mRNA expression in PRNP −/− calves. For protein expression analysis, we performed PrP-specific western blot analyses on fibroblasts (Fig. 1d), peripheral blood lymphocytes (Fig. 1e) and brain stem (Fig. 1f) from wild-type and PRNP −/− calves using the mouse anti-bovine PrP monoclonal antibody F89. We detected PrP-specific bands in the wild-type calves, whereas no reaction was observed in PRNP −/− calves and negative control mouse fibroblasts. These data clearly demonstrate that the PRNP gene is functionally inactivated in the PRNP −/− calves.PRNP −/− cattle were monitored for growth and general health status from birth to 20 months of age. Mean birth weight was 46 kg and average daily gain was 0.91 kg/d to 10 months. Both values were in the normal range for Holstein bulls. Serum chemistry was evaluated at 6 months of age and compared with published reference ranges. All the values for PRNP −/− calves (n = 12) were well within the reference range (Supplementary Table 1) and obvious abnormalities w...

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confidence: 90%

Production of cattle lacking prion protein

et al. 2006

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“…No abnormalities were noted in Prnp °l° mice at the macroscopic, microscopic or behavioral levels [41]. The suggestion that there may be a synaptic deficiency in Prnp °/° mice [42,43] has not been confirmed [44]. The claim that aged mice (with a mixed genetic background) develop ataxia and suffer a loss of Purkinje cells [45] as a consequence of PrP gene disruption is not consistent with previous investigations on independently generated Prnp °/° mouse lines [41,46].…”

Section: Resistance To Scrapie Of Mice Devoid Of Prp Cmentioning

confidence: 99%

“…the events leading to vacuolization and neuronal death are not yet well understood. Because neurons (and astrocytes) may be depleted of PrP c, it has been speculated that this may be the cause of cell damage [42]. The fact that PrP knockout animals show no scrapie-like symptoms and appear quite healthy would argue against this hypothesis, unless one postulates that chronic deprivation of PrP allows for the recruitment of compensatory mechanisms while acute depletion, as may occur in scrapie, does not.…”

Section: Mechanism Of Pathogenesismentioning

confidence: 99%

Molecular biology of transmissible spongiform encephalopathies

1996

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The prion, the transmissible agent that causes spongiform encephalopathies such as serapie, bovine spongiform encephalopathy (BSE) and Creutzfeldt-Jakob disease, is believed to be devoid of nucleic acid and identical with PrP so, a modified form of the normal host protein PrP ¢ which is encoded by the single cop~v gene Prnp. The 'protein only" hypothesis proposes that PrP ~c, when introduced into a normal host, causes the conversion of PrP c into prpS~; it therefore predicts that an animal devoid of PrP c should be resistant to prion diseases. We generated homozygous Prnp °1° ('PrP knockout') mice and showed that, after inoculation with prions, they remained free of scrapie for at least 2 years while wild-type controls all died within 6 months. There was no propagation of prions in the Prnp °t° animals. Surprisingly, heterozygous Prnp °t+ mice, which express PrP c at about half the normal level, also showed enhanced resistance to scrapie disease despite high levels of infectious agent and PrP ~c in the brain early on. After introduction of murine PrP transgenes Prnp °t° mice became highly susceptible to mouse but not to hamster prions, while the insertion of Syrian hamster PrP transgenes rendered them susceptible to hamster but to a much lesser extent to mouse prions. These complementation experiments paved the way to the application of reverse genetics. We have prepared animals transgenic for genes encoding PrP with amino terminal deletions of various lengths and have found that PrP lacking 48 amino proximal amino acids, which comprise four of the five octa repeats of PrP, is still biologically active.

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“…In addition, PrP c , by Cu 2+ binding, may participate in the control of calcium flux and redox stage of the presynaptic terminal (Vassallo and Herms, 2003). Lastly, pioneering electrophysiological studies (using PrP c knockout slices) by Collinge established that PrP c participates in long-term potentiation (Collinge et al, 1994;Curtis et al, 2003;Maglio et al, 2006). More recently, it has been shown that the absence of PrP c alters the expression of several neurotransmitter receptors (see (Maglio et al, 2004;Rangel et al, 2007) for examples).…”

Section: Introductionmentioning

confidence: 99%