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01 Oct 1986TL;DR: Two different cDNAs containing sequences coding for the beta-subunit of bovine follicle stimulating hormone (FSH-beta) have been isolated from a phage lambda gt11 bovina pituitary cDNA library, and the combined sequence represents most of FSH- beta mRNA.
Abstract: Two different cDNAs containing sequences coding for the β-subunit of bovine follicle stimulating hormone (FSH-β) have been isolated from a phage lambda gt11 bovine pituitary cDNA library. The complete nucleotide sequence of both clones was determined, and the combined sequence represents most of FSH-β mRNA. The combined sequence contains 46 nucleotides of 5′-untranslated sequence followed by 387 nucleotides of coding sequence. The coding sequence predicts a 19-amino-acid amino-terminal precursor segment followed by the 110-amino-acid sequence of mature bovine FSH-β. The cDNA sequence demonstrates the presence of a long 3′-untranslated region containing 1295 bases followed by a segment representing the poly(A) portion of the mRNA. Thus, the combined sequence of the cDNAs suggests a minimal size of 1.7 kb for FSH-β mRNA. Analysis of FSH-β sequences present in bovine pituitary mRNA demonstrated the presence of an mRNA with a size of about 2.0 kb. This apparent discrepancy is probably due to the pres...
81 citations
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TL;DR: By fractionation, CFTR was shown to be associated with the membranes that envelop milk fat globules as they are discharged from the apical surface of the mammary epithelia, which represents a novel, inexpensive and efficient approach to produce CFTR and possibly other membrane-associated proteins.
Abstract: Here we describe the production of cystic fibrosis transmembrane conductance regulator (CFTR), the product of the gene associated with cystic fibrosis, in the milk of transgenic mice. Mammary specific expression was achieved by placing the CFTR cDNA under the control of the goat β-casein gene promoter. By fractionation, CFTR was shown to be associated with the membranes that envelop milk fat globules as they are discharged from the apical surface of the mammary epithelia. Since milk fat globules may comprise up to 10% of whole milk, this represents a novel, inexpensive and efficient approach to produce CFTR and possibly other membrane-associated proteins. The availability of large quantities of CFTR could have important implications for the development of new therapies for cystic fibrosis.
81 citations
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TL;DR: It is shown that central nervous system delivery of an adeno-associated viral vector encoding SMN1 produced significant improvements in survival in a mouse model of SMA, and detectable SMN expression in a minimum of 30% motor neurons correlated with efficacy.
Abstract: Spinal muscular atrophy (SMA) is a neuromuscular disease caused by mutations in survival motor neuron 1 (SMN1). Previously, we showed that central nervous system (CNS) delivery of an adeno-associated viral (AAV) vector encoding SMN1 produced significant improvements in survival in a mouse model of SMA. Here, we performed a dose–response study in SMA mice to determine the levels of SMN in the spinal cord necessary for efficacy, and measured the efficiency of motor neuron transduction in the spinal cord after intrathecal delivery in pigs and nonhuman primates (NHPs). CNS injections of 5e10, 1e10, and 1e9 genome copies (gc) of self-complementary AAV9 (scAAV9)–hSMN1 into SMA mice extended their survival from 17 to 153, 70, and 18 days, respectively. Spinal cords treated with 5e10, 1e10, and 1e9 gc showed that 70–170%, 30–100%, and 10–20% of wild-type levels of SMN were attained, respectively. Furthermore, detectable SMN expression in a minimum of 30% motor neurons correlated with efficacy. A comprehe...
81 citations
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TL;DR: Overall, it appears that reports of the death of microscopy have been greatly exaggerated.
81 citations
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TL;DR: The quantitative analysis of mouse ESC-derived neurons and oligodendrocytes co-cultured in a microfluidic device provides insights into the temporal sequence of the myelination process.
Abstract: Myelination in the central nervous system is the process by which oligodendrocytes form myelin sheaths around the axons of neurons. Myelination enables neurons to transmit information more quickly and more efficiently and allows for more complex brain functions; yet, remarkably, the underlying mechanism by which myelination occurs is still not fully understood. A reliable in vitro assay is essential to dissect oligodendrocyte and myelin biology. Hence, we developed a protocol to generate myelinating oligodendrocytes from mouse embryonic stem cells and established a myelin formation assay with embryonic stem cell-derived neurons in microfluidic devices. Myelin formation was quantified using a custom semi-automated method that is suitable for larger scale analysis. Finally, early myelination was followed in real time over several days and the results have led us to propose a new model for myelin formation.
81 citations
Authors
Showing all 3085 results
Name | H-index | Papers | Citations |
---|---|---|---|
George M. Whitesides | 240 | 1739 | 269833 |
Stephen J. O'Brien | 153 | 1062 | 93025 |
Robert B. Jackson | 132 | 458 | 91332 |
Glenn M. Chertow | 128 | 764 | 82401 |
Jon Clardy | 116 | 983 | 56617 |
John J. Fung | 115 | 1011 | 52924 |
Robert B. Colvin | 111 | 556 | 52034 |
Sergio Giralt | 109 | 1024 | 48513 |
Paul Saftig | 107 | 356 | 49929 |
Robert J. Desnick | 102 | 694 | 39698 |
Robert A. Soslow | 87 | 427 | 29014 |
Richard J. Roman | 84 | 461 | 23760 |
Diana W. Bianchi | 81 | 405 | 24554 |
Paolo Raggi | 80 | 439 | 33332 |
Helmut G. Rennke | 77 | 256 | 33959 |