Benjamin J. Reading, Ph.D.

Postdoctoral Research Associate

North Carolina State University
Department of Biology
Campus Box 7617
Raleigh, NC 27695-7617

Office:  377 David Clark Laboratory

Office Phone: (919) 515-3830
Email: bjreadin@unity.ncsu.edu

Email (alt.): ben_reading@unity.ncsu.edu

 


Education

·         Doctor of Philosophy (Zoology), North Carolina State University, Raleigh, NC.  2008.

Dissertation:  Molecular and biochemical characterizations of vitellogenesis in white perch, Morone americana.

·         Master of Science (Zoology), Southern Illinois University, Carbondale, IL.  2003. 

Thesis:  Evidence for polyfactorial sex determination in muskellunge (Esox masquinongy).

·         Bachelor of Arts (Biological Science), Southern Illinois University, Carbondale, IL.  1998.


Professional Experience

 

  • 2008-Present:  Postdoctoral Research Associate, North Carolina State University, Raleigh, NC.
  • 2003-2008:  Graduate Research Assistant, North Carolina State University, Raleigh, NC.
  • 2003:  Logan Hollow Fish Farm, Murphysboro, IL.
  • 1999-2003:  Graduate Research Assistant, Fisheries and Illinois Aquaculture Center, Southern Illinois University, Carbondale, IL.

Interests

 

·         Reproductive Physiology of Fishes, Functional Genomics of Reproduction, Fisheries and Aquaculture.

 

Research

Basic:  Functional genomics of oocyte growth, maturation, and lipid metabolism, with an emphasis on yolk deposition and utilization in fishes.  I have a secondary focus in aquaculture and fisheries management as well. 

Applied:  Indicators of gamete quality and biotechnology related to reproductive dysfunction with an emphasis on manual propagation of cultured fishes and management of fisheries stocks. 

The broad scope of my research emphasis is on reproductive physiology of vertebrates, specifically fishes.  Species important to fisheries and aquaculture, such as striped bass, are particularly suited for this and also serve as valuable model organisms for basic research in biomedicine and agricultural biotechnology.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Manual propagation of fishes.  (Left) Female striped bass broodstock are injected with human chorionic gonadotropin (hCG) to induce ovulation.  Ovulated eggs are then manually stripped from the females and fertilized with sperm from striped bass males.  These fertilized eggs are then incubated in upwelling jars (Right, Top) and once fry hatch from the eggs, reared in aquaria (Right, Bottom) until they can be stocked into outdoor earthen ponds.  

 

I have particular interest in aspects of oogenesis related to deposition and packaging of components in the oocyte that are required for embryonic growth and development.  As examples these include yolk protein precursors, lipids, and egg envelope precursors, and the proteins and mechanisms involved in their production, uptake, processing, and utilization.  To study these processes, I employ a variety of molecular and biochemical techniques to characterize both the gene transcript and protein products and the regulation thereof.

B.J. Reading, Ph.D.

 

Oocyte growth and vitellogenesis in fishes.  (Left) “Oogenesis” is the process by which primordial germ cells (oocytes) become ova (eggs) that are ready to be fertilized.  In fishes and other egg-laying vertebrates, once oogonia become committed to develop as oocytes, they begin to accumulate neutral lipids (during early secondary growth) and then proceed to vitellogenic growth (vitellogenesis), during which time yolk precursors called vitellogenins are taken up and stored within yolk granules in the oocyte.  The yolk serves as a nutrient store for growing embryos and larvae once the eggs have been ovulated and fertilized.  (Right, Top) Induction of vitellogenesis starts with production of follicle stimulating hormone (FSH) in the pituitary.  FSH stimulates production of estradiol (estrogen) in the ovarian follicles, and estrogen in turn induces the liver to produce vitellogenins.  Vitellogenins are released into the blood stream and then specifically incorporated into oocytes by receptor-mediated endocytosis.  (Right, Bottom) Circulating vitellogenins exist as homodimers in the blood.  Once taken up by the oocyte, complete vitellogenins are processed into distinct yolk proteins.  The largest yolk protein is lipovitellin which is important nutritional source of amino acids and polar lipids to support embryonic development.  Phosvitin consists largely of serine residues which act as carriers of phosphorus and calcium and may be involved in the stabilization of vitellogenin structure.  Two more yolk proteins, beta-component and C-terminal component, are known to be released from the C-terminal end of vitellogenin, however, their physiological functions are unknown.  A shorter, incomplete type of vitellogenin also exists that only consists of the lipovitellin yolk proteins.  Vitellogenins are a family of very large particles (400-600kDa in size) and belong to the lipoprotein superfamily, since they consist of both lipid and protein components.  Vitellogenins contain approximately 20% lipid by weight and has been generally believed to be a major transporter of oocyte lipids, the majority of which are structural lipids (polar, phospholipids).  To date, several types of vitellogenins have been characterized in a number of fish species and they have been shown to have specific and distinct nutritive roles during embryogenesis.

 

Central Dogma of Molecular Biology.  Genomics sciences employ a variety of techniques to understand the functions of genes and their related RNA and protein products.

 

Complete Curriculum Vitae [PUT LINK TO CV pdf HERE]


Technical Skills                                                                                               

Biochemistry and Immuno-biochemistry:  Gradient sodium dodecyl-sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) and native  PAGE of proteins; Western blotting (native and SDS-PAGE); protein purification from acrylamide gel matrix; protein purification by column chromatography [open and high performance liquid chromatography (HPLC)] using anion exchange (POROS and MonoQ), gel filtration (Superdex), and affinity (Affigel) media; preparation and absorption of polyclonal antisera (pAbs) for immuno-biochemistry; double immuno-diffusion and Ouchterlony; receptor-ligand blotting; in-well receptor binding assay; digoxigenin (DIG) labeling of proteins; competitive enzyme-linked immunosorbent assay (ELISA); immunocytochemistry.

 

Molecular Biology:  mRNA differential display; Cloning of full-length complementary DNAs (cDNAs) from UniZAPII bacteriophage l cDNA library; in vivo excision of pBluescriptÒ SK(-) phagemid from Uni-ZapTM XR vector using the ExAssistTM/SOLRTM system; 5’- and 3’-rapid amplification of cDNA ends (RACE); nucleic acid probe construction by direct incorporation of DIG and [g-32P]ATP DNA radiolabeling using T4 polynucleotide kinase for DNA-DNA (Southern, colony, and plaque screening) hybridization; reverse transcription (RT) of RNA into cDNA (first strand synthesis); chain termination (dideoxy-nucleotide) DNA sequencing and interpretation; microsatellite isolation from an un-enriched sub-genomic library and microsatellite analyses; AFLP DNA fingerprinting; PAGE and agarose gel electrophoresis (AGE) of nucleic acids; polymerase chain reaction (PCR) and degenerate PCR primer design and optimization of PCR conditions; restriction enzyme digest; molecular cloning of DNA; bacterial transformation, screening, and culture; purification of nucleic acids from tissue (RNA and DNA) and gel (DNA) matrices.

 


Refereed Publications

Reading, B.J., Hiramatsu, N., Sawaguchi, S., Matsubara, T., Hara, A., Lively, M.O., and Sullivan, C.V.  In press.  Conserved and Variant Molecular and Functional Features of Multiple Egg Yolk Precursor Proteins (Vitellogenins) in White Perch (Morone americana) and other Teleosts.  Marine Biotechnology.  [Abstract]

Reading, B.J., Hiramatsu, N., Matsubara, T., Hara, A., and Sullivan, C.V.  2008.  Deduced primary structures of three vitellogenins and specific binding to putative multiple ovarian receptors in white perch (Morone americana).  Cybium International Journal of Ichthyology 32(2) suppl. 159-161.  [Full text]

Davis, L.K., Hiramatsu, N., Hiramatsu, K., Reading, B.J., Matsubara, T., Hara, A., Sullivan, C.V., Pierce, A.L., Hirano, T., and Grau, E.G.  2007.  Induction of three vitellogenins by 17beta-estradiol with concurrent inhibition of the growth hormone-insulin-like growth factor 1 axis in a euryhaline teleost, the tilapia (Oreochromis mossambicus).  Biology of Reproduction 77:614-625.  [Abstract]

Reading, B.J., Wills, P.S., Heidinger, R.C., and Heist, E.J.  2003.  Development of microsatellite markers for muskellunge (Esox masquinongy) and cross-species amplification in two other esocids.  Molecular Ecology Notes 3(3):447-449.  [Abstract]

Heist, E.J., Jenkot, J.L., Keeney, D.B., Lane, R.L., Moyer, G.R., Reading, B.J., and Smith, N.L.  2003.  Isolation and characterization of polymorphic microsatellite loci in nurse shark (Ginglymostoma cirratum).  Molecular Ecology Notes 3(1):59-61.  [Abstract]


Abstracts and Papers Presented at Scientific Meetings and Symposia      

Reading, B.J., Hiramatsu, N., and Sullivan, C.V.  2008.  Reproduction of white perch: multiple egg yolk precursors (vitellogenins) and their receptors.  Invited Seminar, Hormones and the Environment.  Edwin W. Pauley Summer Program in Marine Biology, August 1-15, 2008.  Hawaii Institute of Marine Biology at Coconut Island, Kane’ohe, Hawaii, USA.

Reading, B.J., Hiramatsu, N., Matsubara, T., Hara, A., and Sullivan, C.V.  2007.  Yolk precursors in white perch (Morone americana): deduced primary structures of three types of vitellogenin (Vg) proteins and disparate binding of the different Vgs to multiple ovarian receptors.  Abstract number O 59 In: Abstracts of the 8th International Symposium on Reproductive Physiology of Fish, June 3-8, 2007.  Palais du Grand Large, St. Malo, France.

Reading, B.J., Hiramatsu, N., Williams, V.N., Sullivan, C.V., Sawaguchi, S., Matsubara, T., Amano, and H., Hara, A.  2007.  Multiple egg yolk precursors (vitellogenins) and egg quality in teleost fishes.  Invited Symposium, Physiological Insights Towards Improving Fish Aquaculture.  Abstract number 758 In: Abstracts of Aquaculture America, Aquaculture 2007 Triennial:  Science for Sustainable Aquaculture, February 26-March 2.  San Antonio, Texas, USA.

Sullivan, C.V., Clark, R.W., Kennedy, A.M., Morton, C.L., Hiramatsu, N, Weber, G.M., Klenke, U., and Zohar, Y.  Presented by B.J. Reading.  2004.  Photothermal Control of Final Maturation and Atresia in Temperate Basses (Morone).  Invited Symposium, Nutritional and Physiological Strategies for Sustainable Aquaculture.  Abstract number SO-19-21 In: Abstracts of the 134th Annual Meeting of the American Fisheries Society, August 21-26.  Madison, Wisconsin, USA..

Reading, B.J., Hiramatsu, N., Matsubara, T., Hara, A., and Sullivan, C.V.  2004.  Vitellogenin Multiplicity and Egg Quality in Teleost Fishes.  Invited Symposium, Nutritional and Physiological Strategies for Sustainable Aquaculture.  Abstract number SO-24-05 In: Abstracts of the 134th Annual Meeting of the American Fisheries Society, August 21-26.  Madison, Wisconsin, USA.

Reading, B.J., Wills, P.S., Heidinger, R.C., and Heist, E.J.  2004.  Evidence for a Polyfactorial Sex Determination System in Muskellunge Esox masquinongy. Abstract number CO-174 In: Abstracts of the 134th Annual Meeting of the American Fisheries Society, August 21-26.  Madison, Wisconsin, USA.

Reading, B.J., Wills, P.S., Heidinger, R.C., and Heist, E.J.  2003.  Evidence for polyfactorial sex determination system in muskellunge Esox masquinongy based on gynogenesis, amplified fragment length polymorphism (AFLP) analysis, and microsatellite derived recombination frequencies.  In: Abstracts of the 41st Annual Meeting of the Illinois Chapter of the American Fisheries Society, March 4-6.  Rend Lake State Park, Illinois, USA.  (Lewis L. Osborne Best Student Paper Award).


Professional Presentations and Public Forums                                      

Reading, B.J., Heidinger, R,C., Sheehan, R.J., and Wills, P.S.  2003.  Progress of the Illinois Trophy Muskellunge Project: Sex determination in the muskie and its implications in the production of all female fish.  Annual Meeting of the Shawnee Muskie Hunters Association, March.  Carbondale, Illinois, USA.  

Reading, B.J., Miller-Jackson, A., Heidinger, R.C., Sheehan, R.J., and Wills, P.S.  2001.  Illinois Trophy Muskellunge Project at SIUC.  Annual Meeting of the Shawnee Muskie Hunters Association, March.  Carbondale, Illinois, USA.


Co-Authored Abstracts and Papers           

Sullivan, C.V., Reading, B.J., Hiramatsu, N., Todo, T., Matsubara, T., Sawaguchi, S., Amano, H., and Hara, A.  2008.  Teleost oocyte growth, vitellogenesis, & cytoplasmic maturation.  Invited Plenary Lecture, Sex Determination and Gametogenesis in Fish: Current Status and Future Challenges.  An International Symposium in Honor of Professor Yoshitaka Nagahama, May 30-June 1.  Honolulu, Hawaii, USA. 

Hiramatsu, N., Todo, T., Ito, T., Massaki, K., Kasahara, A., Amano, H., Reading, B.J., Matsubara, T., Sawaguchi, S., Sullivan, C.V., and Hara, A.  2008.  Yolk assembly in teleosts: Recent findings on the deposition of ovarian lipids and proteins.  Invited Symposium, Perspective of Aquaculture Science in Korea and Japan. World Aquaculture 2008: Aquaculture For Human Wellbeing-The Asian Perspective, May 19-23.  Busan, Korea.

Sullivan, C.V., Williams, V.N., Reading, B.J., Hiramatsu, N., Sawaguchi, S., Matsubara, T., Amano, H., and Hara, A.  Presented by C.V. Sullivan.  2006.  The multiple vitellogenin system of Xenotoca eiseni, a moderately matrotrophic Goodeid.  III International Simposium on Viviparous Fishes, Universidad Michoacana de San Nicolás de Hidalgo, Facultad de Biología, November 8-11.  Morelia, Michoacán, México.

Hiramatsu, N., Hiramatsu, K., Reading, B.J., Sullivan, C.V., Sawaguchi, S., Matsubara, T., Amano, H., Hara, A., Weber, G.M., Klenke, U., and Zohar, Y.  Presented by C.V. Sullivan.  2005.  Teleost Oocyte Growth and Ovarian Atresia.  The 1st International Symposium of Potential and Perspective of Marine Bio-manipulation, Enyuu Gakusha Conference Hall, Hokkaido University, February 26-27.  Sapporo, Hokkaido, Japan.


Non-Refereed Publications                                                                                                       

McReynolds, R. and Reading, B.  1998.  DarkTown: The Apocalyptic Cycle.  ISBN: I890305170.  Propaganda Publishing, Cape Girardeau, MO and Gold Rush Games, Elk Grove, CA. 136pp.  Ó1997 by R. McReynolds and B. Reading. 


Teaching Experience                    

 

·         2003-Present:  Supervised several undergraduate and graduate students and trained them in molecular biology, biochemistry, and immuno-biochemistry techniques related to research on the reproductive physiology of fishes. 

·         2005:  Graduate Teaching Assistant, North Carolina State University, Raleigh, NC:  Introduction to Fisheries Science (ZO 420) and Fisheries Science Laboratory (ZO 423).

·         2004:  Graduate Teaching Assistant, North Carolina State University, Raleigh, NC:  Aquaculture I (ZO 586) and Aquaculture I Laboratory (ZO 587).

·         2003:  Graduate Teaching Assistant, North Carolina State University, Raleigh, NC:  Fish Physiology (ZO 515).

·         1999-2002:  Graduate Teaching Assistant, Southern Illinois University, Carbondale, IL:  Comparative Endocrinology (ZOOL426).

·         1999-2001:  Basic Life Support Instructor for the American Heart Association (AHA), Memorial Hospital of Carbondale, Carbondale, IL.

 


Professional Societies and Organizations                                                                                                

 

American Association for the Advancement of Science (AAAS), American Fisheries Society (AFS), American Fisheries Society (AFS) Fish Culture Section, and World Aquaculture Society, US Chapter (WAS).


Professional Services and Activities                                                                                                         

Peer reviewer for the journals Marine Biotechnology (Springer) and North American Journal of Aquaculture (The American Fisheries Society).


Honors and Awards                                                                                                 

Lewis L. Osborne Best Student Paper.  2003.  The 41st Annual Meeting of the Illinois Chapter of the American Fisheries Society, March 4-6, Rend Lake State Park, Illinois (see Abstracts and Papers Presented at Scientific Meetings and Symposia above).

Brevetted Second Lieutenant, Illinois National Guard by Governor Jim Edgar, Executive Office of the Governor of the State of Illinois.  1994.


Current Research Areas

  • Transcriptomics of ovarian genes during the natural reproductive cycle.
  • Follicle atresia and the role of hatching enzymes.
  • Characterization of ovarian lipoprotein receptors and deposition of lipoprotein nutrients in growing oocytes.

B.J. Reading, Ph.D.

 

Hypothetical models proposed for maternal nutrient transfer into Morone oocytes involving several different serum lipoproteins and their related receptor and enzyme systems.  Following ovulation and during embryogenesis, the cargo delivered by LDL and VLDL (non-polar lipids: triacylglycerol, fatty acids, and cholesterol) deposited during oocyte growth contribute largely to egg buoyancy, cell and organelle membrane synthesis, and serve as energy sources during late development, whereas cargo delivered by vitellogenin (polar lipids and protein) contribute to egg buoyancy, protein synthesis, cell and organelle membrane synthesis, and serve as energy sources during early and late embryonic development.  Abbreviations are defined as follows:  A (apolipoprotein A), ApoB/ER (apolipoprotein B and E receptor), ApoER (apolipoprotein E receptor), B (apolipoprotein B), C (apolipoprotein C), CAT (cathepsin), CD36 (fatty acid transporter CD36), CHOL (cholesterol), E (apolipoprotein E), EL (endothelial lipase), FA (fatty acid), FAPB (fatty acid binding protein), HDL (high density lipoprotein), LDL (low-density lipoprotein), LDLR (LDL receptor), LPL (lipoprotein lipase), LR7 (seven ligand binding repeat receptor), LR8 (eight ligand binding repeat receptor), LR8(+) (LR8 receptor containing an O-linked sugar domain), LR8(-) (LR8 receptor lacking an O-linked sugar domain), PL (phospholipid), TAG (triacylglycerol), VLDL (very low-density lipoprotein), VLDLR (VLDL receptor), Vtg (vitellogenin), Vtgr (Vtg receptor). 


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