Genetic Manipulation of the Embryonic Chicken Inner Ear

The mechanisms of formation of the distinct sensory organs of the inner ear and the non-sensory domains that separate them are still unclear. Here, we show that several sensory patches arise by progressive segregation from a common prosensory domain in the embryonic chicken and mouse otocyst. This process is regulated by mutually antagonistic signals: Notch signalling and Lmx1a. Notch-mediated lateral induction promotes prosensory fate. Some of the early Notch-active cells, however, are normally diverted from this fate and increasing lateral induction produces misshapen or fused sensory organs in the chick. Conversely Lmx1a (or cLmx1b in the chick) allows sensory organ segregation by antagonizing lateral induction and promoting commitment to the non-sensory fate. Our findings highlight the dynamic nature of sensory patch formation and the labile character of the sensory-competent progenitors, which could have facilitated the emergence of new inner ear organs and their functional diversification in the course of evolution.

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In vivo genetic manipulation of inner ear connexin expression by bovine adeno-associated viral vectors

Scientific Reports ◽

10.1038/s41598-017-06759-y ◽

2017 ◽

Vol 7 (1) ◽

Cited By ~ 10

Author(s):

Giulia Crispino ◽

Fabian Galindo Ramirez ◽

Matteo Campioni ◽

Veronica Zorzi ◽

Mark Praetorius ◽

...

Keyword(s):

Inner Ear ◽

Viral Vectors ◽

Genetic Manipulation ◽

Connexin Expression

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Zika virus can directly infect and damage the auditory and vestibular components of the embryonic chicken inner ear

Developmental Dynamics ◽

10.1002/dvdy.176 ◽

2020 ◽

Vol 249 (7) ◽

pp. 867-883

Author(s):

Ankita Thawani ◽

Nabilah H. Sammudin ◽

Hannah S. Reygaerts ◽

Alexis N. Wozniak ◽

Vidhya Munnamalai ◽

...

Keyword(s):

Inner Ear ◽

Zika Virus ◽

Embryonic Chicken

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Artificial Induction of Sox21 Regulates Sensory Cell Formation in the Embryonic Chicken Inner Ear

PLoS ONE ◽

10.1371/journal.pone.0046387 ◽

2012 ◽

Vol 7 (10) ◽

pp. e46387 ◽

Cited By ~ 12

Author(s):

Stephen D. Freeman ◽

Nicolas Daudet

Keyword(s):

Inner Ear ◽

Sensory Cell ◽

Cell Formation ◽

Embryonic Chicken ◽

Artificial Induction

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Otoconia formation in the chick embryo

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100076494 ◽

1983 ◽

Vol 41 ◽

pp. 572-573

Author(s):

C.D. Fermin ◽

M. Igarashi

Keyword(s):

Chick Embryo ◽

Inner Ear ◽

Gallus Domesticus ◽

The Body ◽

Abnormal Behavior ◽

Intensive Study ◽

Basic Fuchsin ◽

Gestational Period ◽

Sensory Epithelia ◽

Transmission Electron

Otoconia are microscopic geometric structures that cover the sensory epithelia of the utricle and saccule (gravitational receptors) of mammals, and the lagena macula of birds. The importance of otoconia for maintanance of the body balance is evidenced by the abnormal behavior of species with genetic defects of otolith. Although a few reports have dealt with otoconia formation, some basic questions remain unanswered. The chick embryo is desirable for studying otoconial formation because its inner ear structures are easily accessible, and its gestational period is short (21 days of incubation).The results described here are part of an intensive study intended to examine the morphogenesis of the otoconia in the chick embryo (Gallus- domesticus) inner ear. We used chick embryos from the 4th day of incubation until hatching, and examined the specimens with light (LM) and transmission electron microscopy (TEM). The embryos were decapitated, and fixed by immersion with 3% cold glutaraldehyde. The ears and their parts were dissected out under the microscope; no decalcification was used. For LM, the ears were embedded in JB-4 plastic, cut serially at 5 micra and stained with 0.2% toluidine blue and 0.1% basic fuchsin in 25% alcohol.

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Metabolic engineering of CHO cells to prepare glycoproteins

Emerging Topics in Life Sciences ◽

10.1042/etls20180056 ◽

2018 ◽

Vol 2 (3) ◽

pp. 433-442 ◽

Cited By ~ 1

Author(s):

Qiong Wang ◽

Michael J. Betenbaugh

Keyword(s):

Metabolic Engineering ◽

Cho Cells ◽

Genetic Manipulation ◽

Chinese Hamster ◽

Post Translational Modification ◽

Effector Functions ◽

Recombinant Glycoproteins ◽

Production Platform ◽

Chemical Inhibitors ◽

Amino Acid Mutations

As a complex and common post-translational modification, N-linked glycosylation affects a recombinant glycoprotein's biological activity and efficacy. For example, the α1,6-fucosylation significantly affects antibody-dependent cellular cytotoxicity and α2,6-sialylation is critical for antibody anti-inflammatory activity. Terminal sialylation is important for a glycoprotein's circulatory half-life. Chinese hamster ovary (CHO) cells are currently the predominant recombinant protein production platform, and, in this review, the characteristics of CHO glycosylation are summarized. Moreover, recent and current metabolic engineering strategies for tailoring glycoprotein fucosylation and sialylation in CHO cells, intensely investigated in the past decades, are described. One approach for reducing α1,6-fucosylation is through inhibiting fucosyltransferase (FUT8) expression by knockdown and knockout methods. Another approach to modulate fucosylation is through inhibition of multiple genes in the fucosylation biosynthesis pathway or through chemical inhibitors. To modulate antibody sialylation of the fragment crystallizable region, expressions of sialyltransferase and galactotransferase individually or together with amino acid mutations can affect antibody glycoforms and further influence antibody effector functions. The inhibition of sialidase expression and chemical supplementations are also effective and complementary approaches to improve the sialylation levels on recombinant glycoproteins. The engineering of CHO cells or protein sequence to control glycoforms to produce more homogenous glycans is an emerging topic. For modulating the glycosylation metabolic pathways, the interplay of multiple glyco-gene knockouts and knockins and the combination of multiple approaches, including genetic manipulation, protein engineering and chemical supplementation, are detailed in order to achieve specific glycan profiles on recombinant glycoproteins for superior biological function and effectiveness.

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